Promoters Used to Regulate Gene Expression - Overview
Regulatory elements are crucial to gene expression. This paper discusses the
patent landscape of some widely used transcriptional regulators that are
constitutively active, spatially active (e.g. tissue-specific or
tissue-preferred), and temporally active (e.g. induced or active in
response to a certain chemical or physical stimulus) in plants.
About this technology landscape
- This landscape was originally authored by Carolina Roa-Rodríguez with
funding from the Rockefeller
Foundation. Except where a more recent date is noted on a page, text below
was published in October 2003. Some patents listed in 2003 have lapsed, and new
ones have issued.
- Certain sections of this landscape were updated in 2005-2006 by Dr. Wei Yang
and Dr. Marie Connett Porceddu. Dates are shown on the pages that were updated.
- We are grateful for legal input by Dr. Dianne Rees and for technical
assistance from Dr Nick dos Remedios, Steve Irwin, and Annet Maurer for the web
version.
Chapter 1
Introduction
When the first draft of the human genome sequence became available in
February 2001 there was some surprise that instead of 100,000 genes, only about
30,000 genes were counted. Whole genome sequencing has contributed to the
demise of the paradigm that a single gene or at most a few of them encode each
character or attribute. On the other hand, there's ever more evidence that gene
regulation is one of the major sources of diversity in the phenotypes seen in
nature. Although all cells of one organism contain more or less the same genetic
information, some genes are turned on and others are turned off at different
locations and times during the life cycle of an organism.
Cis- and trans-acting factors regulate gene expression
The intricate pattern of gene regulation involves molecular signals that act
on DNA sequences encoding protein products. Cis-acting molecules act
upon and modulate the expression of physically adjacent, operably linked
polypeptide-encoding sequences. Trans-acting factors affect the
expression of genes that may be physically located very far away, even on
different chromosomes. The expression of a particular gene may be regulated by
the concerted action of both cis and trans-acting elements.
What is the role of promoters in the regulation network?
The promoters discussed throughout the paper are DNA sequences that bind to
the RNA polymerase II enzyme, which is responsible for the generation of RNA.
The promoter region is usually assumed to be the key cis-acting
regulatory region that controls the transcription of adjacent coding region(s)
into messenger ribonucleic acid (mRNA), which is then directly translated into
proteins. DNA sequences within promoters can be identified as binding sites
for trans-acting factors, "transcription factors", which may cause
activation or repression of transcription.
Types of promoters used to regulate gene expression
Promoters used in biotechnology are of different types according to the
intended type of control of gene expression. They can be generally divided into:
-
Constitutive promoters. These promoters direct expression
in virtually all tissues and are largely, if not entirely, independent of
environmental and developmental factors. As their expression is normally not
conditioned by endogenous factors, constitutive promoters are usually active
across species and even across kingdoms.
-
Tissue-specific or development-stage-specific promoters.
These direct the expression of a gene in specific tissue(s) or at certain stages
of development. For plants, promoter elements that are expressed or affect the
expression of genes in the vascular system, photosynthetic tissues, tubers,
roots and other vegetative organs, or seeds and other reproductive organs can be
found in heterologous systems (e.g. distantly related species or even
other kingdoms) but the most specificity is generally achieved with homologous
promoters (i.e. from the same species, genus or family). This is
probably because the coordinate expression of transcription factors is necessary
for regulation of the promoter's activity.
-
Inducible promoters. Their performance is not conditioned
to endogenous factors but to environmental conditions and external stimuli that
can be artificially controlled. Within this group, there are promoters modulated
by abiotic factors such as light, oxygen levels, heat, cold and wounding. Since
some of these factors are difficult to control outside an experimental setting,
promoters that respond to chemical compounds, not found naturally in the
organism of interest, are of particular interest. Along those lines, promoters
that respond to antibiotics, copper, alcohol, steroids, and herbicides, among
other compounds, have been adapted and refined to allow the induction of gene
activity at will and independently of other biotic or abiotic factors.
-
Synthetic promoters. Promoters made by bringing together
the primary elements of a promoter region from diverse origins.
Apart from the promoter types mentioned above, there are regulatory
expression systems based on transactivating proteins. These systems regulate the
expression of genes of interest irrespective of their physical position to the
target genes. In fact, several chemical-inducible promoters incorporate
transactivating proteins and constitutive promoters as part of the regulatory
system. Transactivating proteins constitute a whole realm of molecules in the
field of gene regulation and deserve a separate analysis. We deal with some of
them in this paper, but only to the extent that they overlap with the types of
promoters aforementioned.
Why the interest in promoters?
The interest in promoters stems from the myriad opportunities for controlling
gene expression. The study and understanding of the function of their multiple
components and the factors associated with their performance have opened up the
possibility of modulation of the expression of genes in homologous organisms as
well as in heterologous organisms, where foreign promoters together with genes
of interest are inserted. Promoters are regarded as molecular biological tools
crucial for the regulation of the expression of genes of interest. As
such, promoter-related patents have a huge influence in follow-on research and
development in biotechnology.
What sort of information do we provide on promoters?
There is virtually an endless number of promoters, potentially as many as
there are genes. For example, a diploid flowering plant has an estimated 25 000
genes. As full genome sequences of different organisms (e.g., such as
Arabidopsis) are becoming available, a great number of promoters are
being identified, isolated and evaluated, and many more are likely to pop up in
the near future. If we attempted to present them all, it would be a daunting and
almost never-ending task. Our aim instead is to present scientific and patent
information on some of the most widely used promoters.
The information included in this document comprises:
-
Primary information about promoters. Over the last two
decades, the concept of what a promoter is has changed. Previously considered
to be a string of a few functional sequence elements, promoter sequences are
now known to include many structural elements which can have complex
functional interactions. We present a view of a consensus promoter, its common
elements and functions and describe the basic differences between eukaryotic
promoters and prokaryotic promoters. This information has been compiled from a
variety of scientific books and journal articles.
-
Types of promoters. This section describes different
categories of promoters: constitutive promoters, tissue-specific promoters,
inducible promoters, and synthetic promoters and their modes of operation,
properties, applications and drawbacks. A selection of widely used promoters
within these categories is presented in detail. The scientific information has
been compiled from scientific books and printed and on-line journals, and
information provided by researchers.
-
Intellectual property aspects of promoters. As in many
burgeoning areas of science with potential far-reaching commercial applications,
promoters and their uses are being patented. The analysis of every promoter
subject to patent protection would be overly time consuming and probably
ultimately not very useful, because almost every gene is likely to be a source
of a promoter. Instead, our analyses focus on patents with claims relating to
the broad categories of promoters described above, and on details of selected
patents with claims relating to specific types of promoters, in some cases. The
information deals with:
-
Patents on selected widely-used promoters and general promoter
types. We assess the possibilities for, and limitations on, further
development of technologies that are based on understanding controls that
regulate gene expression. The patent analyses provided include general patent
information such as patent numbers, total number of patents identified relating
to a particular promoter, applicant names, dates of filing, priority claims,
grant dates and expected expiry dates. Additionally, we analyse claims of
selected relevant patents, including aspects of the prosecution history
of certain patents.
-
Overlapping patent protection. Although the inventions
protected by individual patents cannot be exactly the same, in certain cases,
there are patents that due to the breadth of their claim scope may encompass
other protected inventions. Additionally, there may be patents sharing common
features. Where this is the case, this report attempts to point out
the relationships between claims of different patents and the possible room left
to maneuver around these claims. Take into account that there may be patents
that have not been identified in our searches and further, that there may
be patents, that while not entirely about promoters, may have an effect on
limiting freedom to operate in technologies relating to regulation of gene
expression. For example, this is the case for the restrictive reproductive
technologies, dubbed "Terminator" and " Verminator" technologies, which may
have a great impact on the use and development of methods to regulate the
expression of genes related to plant reproduction and seed generation.
-
IP rights and their impact on the commercialization of technology
based on promoters. Although many people regard studies of promoters as
being confined to academic research circles, press releases and information
provided by agrobiotech companies illustrate how patent rights are used as
commercial assets in the agrobiotechnology industry. Patents with claims
relating to plant regulatory regions may circumscribe subsequent development and
innovation in research on promoters and other research areas that rely heavily
on understanding mechanisms for controlling gene expression.
Primary information about promoters
Eukaryotic promoters
To turn a gene into a protein product, at least two general steps are
required:
- the gene is transcribed, spliced and processed to form mRNA, and
- the mRNA is translated into a polypeptide.
Transcription is a controlled process. While multiple DNA regions are
involved, the promoter is the main determinant for the initiation of
transcription and modulation of levels and timing of gene expression.
Promoters in eukaryotic organisms- e.g. plants, animals- comprise multiple
elements, some of which are found in nearly all promoters. These include:
-
CAAT box. A consensus sequence close to -80 bp from the
start point (+1). It plays an important role in promoter efficiency, by
increasing its strength, and it seems to function in either orientation. This
box is replaced in plants by a consensus sequence called the AGGA
box;
-
TATA box. A sequence usually located around 25 bp upstream
of the start point. The TATA box tends to be surrounded by GC rich sequences.
The TATA box binds RNA polymerase II and a series of transcription factors
(TFIIX, X being a letter that identifies an individual transcription factor) to
form an initiation complex;
-
GC box. A sequence rich in guanidine (G) and cytidine (C)
nucleotides, is usually found in multiple copies in the promoter region,
normally surrounding the TATA box; and
-
CAP site. A transcription initiation sequence or start
point defined as +1, at which the transcription process actually starts.
| Conserved eukaryotic promoter elements |
Consensus sequence |
|
CAAT box
|
GGCCAATCT
|
|
TATA box
|
TATAA
|
|
GC box
|
GGGCGG
|
|
CAP site
|
TAC
|
RNA polymerase II is the enzyme that transcribes a gene into RNA. It works in
conjunction with other transcription factors that recognize signals embodied in
the promoter region. RNA polymerase II starts its "journey" at the TATA region
where it binds and travels along the DNA until it reaches the CAP site where the
actual synthesis of RNA starts. The transcription process only takes place in
the downstream direction, from 5' (left) to 3' (right).
These elements are normally regarded as constituents of the
promoter region itself, but depending on the scope of definition of a promoter
in a patent or patent application, and whether the definition is expressed in
functional or structural terms, other elements may be included as part of a
promoter region.
Enhancers, for example, are elements located at variable distances from the
promoter "itself" and contain several closely arranged sequence elements that
bind to transcription factors. These elements enhance the activity of a promoter
and are orientation-independent with respect to the promoter and can be upstream
or downstream of a promoter (e.g., such as within intron sequences of a gene).
There is currently a high interest in studying and isolating enhancers, which
can be successfully attached to heterologous promoter regions to increase
transcriptional activity and in some cases to provide additional levels of
control (e.g., to confer tissue-specific or stage-specific expression of a
gene).
Prokaryotic promoters
Promoters of prokaryotic organisms, e.g., such as bacteria, have similar
elements as the eukaryotic promoters although there are a few basic
differences. Prokaryotic promoters contain at least three conserved features
defining the region where the RNA polymerase binds:
- the start point, defined as +1;
- the TATA box is located at -10 position to the start point;
in contrast to the -35 bp in eukaryotic promoters; and
- the TTGACA sequence, also called the -35
sequence, located around 35 bp upstream of the start point.
An additional feature, much more common in prokaryotic organisms, is that a
promoter serves to initiate the transcription of multiple structural
genes that are immediately adjacent to it. This arrangement is called
an operon. A single transcribed mRNA is translated into several
proteins whose functions are interrelated. In operons, promoters have adjacent,
juxtaposed or interspersed regulatory sites to which regulatory proteins bind.
In eukaryotic promoters, the regulatory sites are spread out over a longer
distance.
There are two modes of regulation of the initiation of transcription in
operons:
-
Positive control mode, where the interaction between the
regulatory protein and regulatory region on the DNA turn the transcription on.
The genes are off by default and are turned on by the
activators. Transcription factors interact with the RNA
polymerase and assist the enzyme in initiating transcription at the promoter.
This positive fashion of controlling gene expression is more common in
eukaryotes than in prokaryotes.
-
Negative control mode, where the interaction turns the
genes off. In this case, a repressor protein binds the
operator, a DNA sequence of approximately 20 to 25 nucleotides,
which is next to the promoter or juxtaposed, and prevents the RNA polymerase
from initiating transcription. To switch on the system, small molecules called
inducers trigger the production of proteins by binding to the
repressor protein and changing its conformation. This change alters the
operator-repressor interaction, so that the repressor can no longer remain
attached to the operator. Negative control is widely used among prokaryotes,
which need to respond swiftly to changes in the environment.
One of the best-studied operon systems is the lac operon from
Escherichia coli (E. coli). Since its discovery in the 1960s,
other operon systems have been extensively studied in other organisms and are
currently being adapted to plant systems with the aim of tightly regulating the
expression of genes in transgenic plants.
Promoter types
Summary
As mentioned in the introduction, promoters can be categorized according to
the type or degree of control of gene expression: control in all or virtually
all tissues or control depending on the tissue and the developmental stage of
the plant. Additionally, promoters may operate in response to external and, in
some cases, controllable stimuli. Thus, they can be classified as follows:
-
Constitutive promoters, which
induce the expression of the downstream-located coding region in all tissues
irrespective of environmental or developmental factors.
-
Synthetic promoters, which comprise
consensus DNA sequences of common elements of natural promoter regions.
-
Inducible promoters, which are only
expressed under the presence of factors/compounds. Because their expression is
normally restricted to certain plant tissues, they can also be considered as
tissue-specific. Based on the nature of the factors that trigger their
expression, they are divided into two groups:
-
Chemically-regulated, where
chemical compounds, usually not naturally found within plants, switch on
promoter activity. Several of the types of promoters discussed in this paper
involve chimeric components gathered from human, animal, fungal and bacterial
sources.
-
Physically-regulated, where abiotic
and external factors such as light, heat, mechanical injury induce promoter
activity.
-
Tissue-specific promoters, which
operate in particular tissues and at certain developmental stages of a plant.
They may be induced by endogenous and exogenous factors, so they may be also
classified as inducible.
Chapter 2 - Constitutive promoters
Summary
There are several advantages to using constitutive promoters in expression
vectors used in plant biotechnology, such as:
- High level of production of proteins used to select transgenic cells or
plants;
- High level of expression of reporter proteins or scorable markers, allowing
easy detection and quantification;
- High level of production of a transcription factor that is part of a
regulatory transcription system;
- Production of compounds that requires ubiquitous activity in the plant; and
- Production of compounds that are required during all stages of plant
development
The first constitutive promoters used for the expression of transgenes in
plants were isolated from plant pathogens.
The search for other constitutive promoters has continued, especially to
identify control regions that are able to drive expression of transgenes in
monocots. In some monocots such as cereals, it has been found that sequences
present in 5' untranslated transcribed regions (e.g., introns) of certain
structural genes are essential for efficient gene expression. Thus, promoters
that work well in dicots, which lack introns, do not generally work well in
monocots. For this reason, regulatory sequences from monocots have also
been identified and cloned into vectors for control of transgene expression. The
following constitutive promoters are among the most widely used nowadays and are
the ones analyzed in this report.
-
Plant pathogen promoters
-
Monocot promoters
The CaMV 35S promoter
Analysis on the CaMV 35 promoter is divided into a discussion of:
- the promoter itself
- sequences identified in patents as "35S enhancer regions"
- the "minimal" promoter
Scientific aspects
At the beginning of the 1980s, Chua and collaborators at the Rockefeller
University isolated the promoter responsible for the transcription of the whole
genome of a Cauliflower mosaic virus (CaMV) infecting turnips. The promoter was
named CaMV 35S promoter ("35S promoter")
because the coefficient of sedimentation of the viral transcript whose
expression is naturally driven by this promoter is 35S. It is one of the most
widely used, general-purpose constitutive promoters.
The 35S promoter is a very strong constitutive promoter, causing high levels
of gene expression in dicot plants. However, it is less effective in monocots,
especially in cereals. The differences in behavior are probably due to
differences in quality and/or quantity of regulatory factors.
The promoter responsible for the transcription of another part of the genome
of CaMV, the CaMV 19S promoter, is also used as a constitutive
promoter, but is not as widely used as the 35S promoter.
CaMV 35S promoter IP issues
Monsanto Company and The
Rockefeller University are the owners of patents on the CaMV
35S promoter. The geographical range of their patents, according to publicly
available information sources, is limited to:
- The United States and Japan in the case of Monsanto (a patent may still be
pending in Brazil), and
- Only the United States in the case of the Rockefeller University.
If you are aware of additional or more up to date information about patent
coverage, please add it to what is available to users through our comments
interface.
How is promoter defined?
Before entering the discussion about claims granted, it is important to know
how a promoter is defined in the patents.
Monsanto defines promoter in a functional manner. In the
Monsanto patents a promoter is the region at the 5' end of a gene that initiates
transcription of the gene to produce a mRNA transcript. The 35S promoter in
particular is referred to as the promoter for the full-length
mRNA of the CaMV genome.
The Rockefeller University does not explicitly
define the word "promoter." Nevertheless, in the specifications of the US
patents, a promoter sequence is described as necessary in
order to obtain "adequate expression" of a gene inserted into a plant. So, in
that sense, the concept of a promoter is also functional. With
respect to the 35S promoter, the specifications disclose the use of CaMV 35S
promoter as a promoter for general use. They define the 35S
promoter in terms of the sequences of its subdomains.
Approximate scope of claims
Both entities have claims directed to methods and products, but they address
different aspects of the 35S promoter. In general terms,
- Monsanto's patents are directed to chimeric genes containing the 35S or the
19S promoter controlling a heterologous gene, and
- The Rockefeller University's patents are directed to the DNA sequences of
the individual subdomains of the 35S promoter, combinations of them, and the use
of B subdomains in particular to form tissue-specific promoters.
In the U.S., there are currently four patents granted to Monsanto and two
patents granted to The Rockefeller University. This situation may change over
time as additional applications may be pending and some may lapse; if you are
aware of more current information, please add comments on this page or on the
page giving details on the patents and claims.
The U.S. claims cover:
- Monsanto
- "Chimeric genes" having the CaMV 35S promoter or the CaMV 19S
promoter, a heterologous gene and a poly(A) signal;
- In particular, such genes in which the heterologous gene confers antibiotic
resistance to a transformed plant;
- Plant transformation vectors, both intermediates and vectors having a
chimeric gene as described;
- Differentiated dicot plants containing these chimeric genes; and
- Methods for transforming plant cells with these chimeric genes.
- The Rockefeller University
- Isolated DNA sequences of four of the five subdomains of domain B of the
CaMV 35S promoter;
- The same sequences of the B subdomains linked to domain A of the 35S
promoter and to the minimal promoter region of the 35S promoter;
- Sequence of a tissue-specific promoter corresponding to domain B of the 35S
promoter;
- Method for the tissue-specific expression of a chimeric gene by using domain
B coupled to the domain A of the 35S promoter.
Claims in the European patent granted to Monsanto covered a "chimeric gene"
very similar to the one claimed in the U.S., except that the promoter is limited
to the 35S promoter and the chimeric gene also contains a 5' non-translated
region. This patent has now lapsed, but it is worth mentioning because the
European patent as first granted was opposed by several institutions. The claim,
in the application as originally filed, was directed to a promoter from a plant
virus, which could be from any plant virus, not only from CaMV. After the
opposition, the promoter was restricted to the CaMV 35S promoter.
The specific patent information and a summary of the independent claims of
each patent are presented in the following tables.
Patents on 35S promoter
Patent Number
|
Title, Independent Claims and Summary of Claims
|
Assignee
|
|
US
5352605
- Earliest priority - 17 January 1983
- Filed - 28 October 1993
- Granted - 4 October 1994
- Expected expiry - 4 October 2011
|
Title - Chimeric genes for transforming plant cells using
viral promoters
Claim 1
A chimeric gene which is expressed in plant cells comprising a promoter from
a cauliflower mosaic virus, said promoter selected from the group consisting of
a CaMV (35S) promoter isolated from CaMV protein-encoding DNA sequences and a
CaMV (19S) promoter isolated from CaMV protein-encoding DNA sequences, and a
structural sequence which is heterologous with respect to the promoter.
|
Claim 4
A plant cell which comprises a chimeric gene that contains a promoter from
cauliflower mosaic virus, said promoter selected from the group consisting of a
CaMV (35S) promoter and a CaMV (19S) promoter, wherein said promoter is isolated
from CaMV protein-encoding DNA sequences, and a structural sequence which is
heterologous with respect to the promoter.
|
Claim 7
An intermediate plant transformation plasmid which comprises a region of
homology to an Agrobacterium tumefaciens vector, a T-DNA border region
from Agrobacterium tumefaciens and a chimeric gene, wherein the
chimeric gene is located between the T-DNA border and the region of homology,
said chimeric gene comprising a promoter from cauliflower mosaic virus, said
promoter selected from the group consisting of a CaMV(35S) promoter and a
CaMV(19S) promoter, and a structural sequence which is heterologous with respect
to the promoter.
|
Claim 8
A plant transformation vector which comprises a disarmed plant tumor inducing
plasmid of Agrobacterium tumefaciens and a chimeric gene, wherein the
chimeric gene contains a promoter from cauliflower mosaic virus, said promoter
selected from the group consisting of a CaMV(35S) promoter and a CaMV(19S)
promoter, and a structural sequence which is heterologous with respect to the
promoter.
|
Claim 13
A DNA construct comprising: (A) a CaMV promoter selected from the group
consisting of (1) a CaMV 35S promoter isolated from CaMV protein-encoding DNA
sequences and (2) a CaMV 19S promoter isolated from CaMV protein-encoding DNA
sequences, and (B) a DNA sequence of interest heterologous to (A), wherein (B)
is under the regulatory control of (A) when said construct is transcribed in a
plant cell.
|
Claim 14
A chimeric gene which is transcribed and translated in plant cells, said
chimeric gene comprising a promoter from cauliflower mosaic virus, said promoter
selected from the group consisting of: a) a CaMV 35S promoter region free of
CaMV protein-encoding DNA sequences and b) a CaMV 19S promoter region free of
CaMV protein-encoding DNA sequences, and a DNA sequence which is heterologous
with respect to the promoter.
|
Claim 15
A chimeric gene which is expressed in plants cells comprising a promoter from
a cauliflower mosaic virus, said promoter selected from the group consisting of
a CaMV(35S) promoter region free of CaMV protein-encoding DNA sequences and a
CaMV(19S) promoter region free of CaMV protein-encoding DNA sequences, and a DNA
sequence which is heterologous with respect to the promoter.
|
Claim 16
A chimeric gene which is transcribed in plants cells comprising a promoter
from a cauliflower mosaic virus, said promoter selected from the group
consisting of a CaMV(35S) promoter free of CaMV protein-encoding DNA sequences
and a CaMV(19S) promoter free of CaMV protein-encoding DNA sequences, a DNA
sequence which is heterologous with respect to the promoter and a 3'
non-translated polyadenylation signal sequence.
|
Claim 17
A plant cell which comprises a chimeric gene where said chimeric gene
comprises a promoter from cauliflower mosaic virus, said promoter selected from
the group consisting of a CaMV(35S) promoter and a CaMV(19S) promoter, wherein
said promoter is free of CaMV protein-encoding DNA sequences, and a DNA sequence
which is heterologous with respect to the promoter and a 3' non-translated
polyadenylation signal sequence.
|
Claim elements:
- 35S and 19S promoters, as defined in the specification, are
claimed as part of a chimeric construct, also containing a heterologous signal
and a poly(A) signal, expressed in a plant
- There are also claims to intermediate plant transformation vectors and plant
transformation vectors containing the chimeric gene for
Agrobacterium-mediated plant transformation, and to plant cells having
the chimeric gene.
|
Monsanto |
|
US
5530196
- Earliest priority - 17 January 1983
- Filed - 2 September 1994
- Granted - 25 June 1996
- Expected expiry - 25 June 2013
|
Title - Chimeric genes for transforming plant cells using
viral promoters
Claim 1
A differentiated dicotyledonous plant comprising plant cells containing a
chimeric gene which comprises a promoter from cauliflower mosaic virus, said
promoter selected from the group consisting of a CaMV(35S) promoter free of CaMV
protein-encoding DNA sequences and a CaMV(19S) promoter free of protein-encoding
DNA sequences, and a structural sequence which is heterologous with respect to
the promoter.
|
Claim 4
A differentiated dicotyledonous plant comprising plant cells containing in
the plant genome a chimeric gene which comprises a promoter from cauliflower
mosaic virus, said promoter selected from the group consisting of a CaMV(35S)
promoter and a CaMV(19S) promoter, and a DNA sequence which is heterologous with
respect to the promoter.
|
Claim 5
A differentiated dicotyledonous plant regenerated from plant cells, said
plant cells containing a chimeric gene which comprises a promoter from
cauliflower mosaic virus, said promoter selected from the group consisting of a
CaMV(35S) promoter and a CaMV(19S) promoter, and a DNA sequence which is
heterologous with respect to the promoter.
|
This patent is a Continuation of
US
5352605.
- Differentiated dicot plants having plant cells with a
chimeric gene similar as described above. The poly(A) signal is not expressly
claimed as a component of the chimeric gene.
|
|
US
5858742
- Earliest priority - 17 January 1983
- Filed - 24 June 1996
- Granted - 12 January 1999
- Expected expiry - 25 June 2013
|
Title - Chimeric genes for transforming plant cells using
viral promoters
|
Claim 1
A method for transforming a plant cell which
comprises transforming a plant cell with a chimeric DNA construct containing a
promoter isolated from cauliflower mosaic virus (CaMV), said promoter selected
from the group consisting of a CaMV(19S) promoter derived from the CaMV(19S)
gene and a CaMV(35S) promoter derived from the CaMV(35S) gene, and a DNA
sequence which is heterologous with respect to the promoter; wherein the
promoter regulates the transcription of the DNA sequence.
|
This patent is a Continuation of
US
5530196.
- Method for transforming plant cells with a chimeric DNA construct as
generally described in
US
5530196.
|
|
US
6255560
- Earliest priority - 17 January 1983
- Filed - 11 January 1999
- Granted - 3 July 2001
- Expired - 3 August 2005
|
Title - Chimeric genes for transforming plant cells using
viral promoters
|
Claim 1
A chimeric gene which is expressed in plant
cells comprising a promoter from cauliflower mosaic virus (CaMV), wherein said
promoter is the CaMV(19S) promoter or the CaMV(35S) promoter, operably linked to
a DNA sequence which is heterologous with respect to the promoter, wherein: the
promoter regulates the transcription of the DNA sequence, and the DNA sequence
encodes a polypeptide conferring increased antibiotic resistance to a plant or
plant cell containing the DNA sequence relative to a wild-type plant or plant
cell.
|
|
Claim 3
A plant cell comprising a chimeric gene which
comprises a promoter from cauliflower mosaic virus (CaMV), wherein said promoter
is the CaMV(19S) promoter or the CaMV(35S) promoter, operably linked to a DNA
sequence which is heterologous with respect to the promoter, wherein: the
promoter regulates the transcription of the DNA sequence, and the DNA sequence
encodes a polypeptide conferring increased antibiotic resistance to the plant
cell relative to a wild-type plant cell.
|
|
Claim 6
An intermediate plant transformation plasmid
which comprises a region of homology to an Agrobacterium tumefaciens
vector, a T-DNA border from Agrobacterium tumefaciens, and a chimeric gene,
wherein the chimeric gene is located between the T-DNA border and the region of
homology, said chimeric gene comprising a promoter from cauliflower mosaic virus
(CaMV), wherein said promoter is the CaMV(19S) promoter or the CaMV(35S)
promoter, operably linked to a DNA sequence which is heterologous with respect
to the promoter, wherein: the promoter regulates the transcription of the DNA
sequence, and the DNA sequence encodes a polypeptide conferring increased
antibiotic resistance to a plant or plant cell containing the DNA sequence
relative to a wild-type plant or plant cell.
|
|
Claim 9
A plant transformation vector which comprises a
modified plant tumor inducing plasmid of Agrobacterium tumefaciens
which is capable of inserting a chimeric gene into susceptible plant cells,
wherein the chimeric gene comprises a promoter from cauliflower mosaic virus
(CaMV), wherein said promoter is the CaMV(19S) promoter or the CaMV(35S)
promoter, operably linked to a DNA sequence which is heterologous with respect
to the promoter, wherein: the promoter regulates the transcription of the DNA
sequence, and the DNA sequence encodes a polypeptide conferring increased
antibiotic resistance to a plant or plant cell containing the DNA sequence
relative to a wild-type plant or plant cell.
|
|
Claim 12
A differentiated dicotyledonous plant
comprising plant cells containing a chimeric gene which comprises a promoter
from cauliflower mosaic virus (CaMV), wherein said promoter is the CaMV(19S)
promoter or the CaMV(35S) promoter, operably linked to a DNA sequence encoding
said polypeptide which is heterologous with respect to the promoter, wherein:
the promoter regulates the transcription of the DNA sequence, and the DNA
sequence encodes a polypeptide conferring increased antibiotic resistance to the
plant relative to a wild-type plant.
|
This patent is a Continuation of
US
5858742.
- A chimeric gene where the 35S or the 19S promoter regulates the
transcription of a gene conferring antibiotic resistance to a plant.
- An intermediate plant transformation vector with the mentioned chimeric gene
between a T-DNA border and a region of homology to an Agrobacterium tumefaciens
vector.
- A plant transformation vector capable of inserting the chimeric gene into a
plant cells. Differentiated dicot plants having plant cells with the chimeric
gene.
|
|
EP
131623 B2
- Earliest priority - 17 January 1983
- Filed - 16 January 1984
- Granted - 28 July 1999
- Lapsed after termination of 20 years
|
Title - Chimeric genes suitable for expression in plant
cells
|
Claim1
A chimeric gene capable of expressing a neomycin
phospotransferase polypeptide in plant cells conferring antibiotic resistance to
the plant when inserted into the plant genome, comprising in sequence:
a) a promoter region from a ribulose-1,5-bis-phosphate carboxyase small
subunit gene;
b) a 5' non-translated region
c) a structural coding
sequence encoding neomycin phospotransferase I or II; and
d) a 3'
non-translated region of a gene naturally expressed in plant cells, said region
encoding a signal sequence for polyadenylation of mRNA, said promoter region
being heterologous with respect to the structural coding sequence .
|
|
Claim 4
A chimeric gene capable of expressing a
polypeptide in plant cells comprising in sequence:
a) a full-length transcript promoter region isolated from cauliflower mosaic
virus
b) a 5' non-translated region
c) a structural coding
sequence
d) a 3' non-translated region of a gene naturally expressed in
plants, said region encoding a signal sequence for polyadenylation of mRNA, said
structural coding sequence being heterologous with respect to said promoter
region.
|
|
Claim 6
A culture of microorganisms identified by ATCC
accession number 39265.
|
The claims are to:
- A chimeric gene capable of expressing a polypeptide in plant cells having a
35S promoter, an intron, a structural coding sequence and a
poly(A) signal.
|
|
Remarks
|
Granted Japanese patent JP 2645217 was not analyzed. There
are also a pending Japanese application (JP 7014349 B4) and a
pending Brazilian application (BR 1101069).
|
|
Note: Patent information on this page was last updated on 15 March 2006.
Patents on subdomains of the 35S promoter
|
Patent number
|
Title, Summary of Claims and Independent Claims
|
Assignee
|
|
US
5097025
- Earliest priority - 1 August 1989
- Filed - 1 December 1989
- Granted - 12 May 1992
- Expected expiry - 1 August 2009
|
Title - Plant Promoters
Claim elements:
- Isolated DNA sequences of four of the five subdomains of domain B of the 35S
promoter (see
diagram).
- DNA sequences of the combinations of each of the four subdomains B and the
domain A of the 35S promoter.
- DNA sequences of the combinations of each of the four subdomains B and the
minimal promoter of the 35S promoter.
|
Claim 1
An isolated DNA segment consisting of the nucleotide sequence:
5'-CGACCAGCAT CGTGGAAAAA GAAGACGTTC CAACCACGTC TTCAAAGC-3'*.
*sequence of subdomain B2 of 35S CaMV promoter.
|
|
Claim 2
A DNA sequence consisting of:
a) a first nucleotide sequence 5'-CGAGGAGCAT CGTGGAAAAA GAAGACGTTC CAACCACGTC
TTCAAAGC-3'* and
b) a second nucleotide sequence corresponding to domain A of the CaMV 35S
promoter coupled to said first nucleotide sequence by means of a synthetic
multilinker.
*sequence of subdomain B2 of 35S CaMV promoter.
|
|
Claim 3
A DNA sequence consisting of:
a) a first nucleotide sequence 5'-CATCGTTGAAG ATGCCTCTGC CGACAGTGGT CCCAAAGATG
GACCCCCACC CAC-3'* and
b) a second nucleotide sequence corresponding to domain A of the CaMV35S
promoter coupled to said first nucleotide sequence by means of a synthetic
multilinker.
*sequence of subdomain B3 of 35S CaMV promoter.
|
|
Claim 4
A DNA sequence consisting of:
a) a first nucleotide sequence 5'-ATTCC ATTGCCC AGCTATCTGT CACTTTATTG TGAAGATAGT
GGAAAAGGAA GGTGGCTCCT ACAAATGCCA TCATTGCGAT AAAGGAAAGG CC-3'* and
b) a second nucleotide sequence corresponding to domain A of the CaMV 35S
promoter coupled to said first nucleotide sequence by means of a synthetic
multilinker.
*sequence of subdomain B4 of 35S CaMV promoter.
|
|
Claim 5
A DNA sequence consisting of:
a) a first nucleotide sequence 5'-TGAGACTTTT CAACAAAGGG TAATATCCGG AAACCTCCTC
GGATT-3'* and
b) a second nucleotide sequence corresponding to domain A of the CaMV 35S
promoter coupled to said first nucleotide sequence by means of a synthetic
multilinker.
*sequence of subdomain B5 of 35S CaMV 35S promoter.
|
|
Claim 6
An isolated DNA segment consisting of the nucleotide sequence:
5'-CATCGTTGAAG ATGCCTCTGC CGACAGTGGT CCCAAAGATG GACCCCCACC CAC-3'*.
*sequence of subdomain B3 of 35S CaMV promoter.
|
|
Claim 7
An isolated DNA segment consisting of the nucleotide sequence:
5'-ATTCC ATTGCCC AGCTATCTGT CACTTTATTG TGAAGATAGT GGAAAAGGAA GGTGGCTCCT
ACAAATGCCA TCATTGCGAT AAAGGAAAGG CC-3'*.
*sequence of subdomain B4 of 35S CaMV promoter.
|
|
Claim 8
An isolated DNA segment consisting of the nucleotide sequence:
5'-TGAGACTTTT CAACAAAGGG TAATATCCGG AAACCTCCTC GGATT-3'*.
*sequence of subdomain B5 of 35S CaMV promoter.
|
|
Claim 9
A DNA sequence consisting of:
a) a first nucleotide sequence 5'-CGAGGAGCAT CGTGGAAAAA GAAGACGTTC CAACCACGTC
TTCAAAGC-3'* and
b) a second nucleotide sequence corresponding to the minimal promoter region of
the CaMV 35S promoter coupled to said first nucleotide sequence by means of a
synthetic multilinker.
*sequence of subdomain B2 of 35S CaMV promoter.
|
|
Claim 10
A DNA sequence consisting of:
a) a first nucleotide sequence 5'-CATCGTTGAAG ATGCCTCTGC CGACAGTGGT CCCAAAGATG
GACCCCCACC CAC-3'*
and
b) a second nucleotide sequence corresponding to the minimal promoter region of
the CaMV 35S promoter coupled to said first nucleotide sequence by means of a s
ynthetic multilinker.
*sequence of subdomain B3 of CaMV 35S promoter
|
|
Claim 11
A DNA sequence consisting of:
a) a first nucleotide sequence 5'-ATTCC ATTGCCC AGCTATCTGT CACTTTATTG TGAAGATAGT
GGAAAAGGAA GGTGGCTCCT ACAAATGCCA TCATTGCGAT AAAGGAAAGG CC-3'* and
b) a second nucleotide sequence corresponding to the minimal promoter region of
the CaMV 35S promoter coupled to said first nucleotide sequence by means of a
synthetic multilinker.
*sequence of subdomain B4 of 35S CaMV promoter
|
|
Claim 12
A DNA sequence consisting of:
a) a first nucleotide sequence 5'-TGAGACTTTT CAACAAAGGG TAATATCCGG AAACCTCCTC
GGATT-3'* and
b) a second nucleotide sequence corresponding to the minimal promoter region of
the CaMV 35S promoter coupled to said first nucleotide sequence by means of a
synthetic multilinker.
*sequence of subdomain B5 of 35S CaMV promoter
|
|
The Rockefeller University
|
|
US
5110732
- Earliest priority - 14 March 1989
- Filed - 14 March 1989
- Granted - 5 May 1992
- Expected expiry - 5 May 2009
|
Title - Selective gene expression in plants
Claim elements:
- a tissue-specific promoter fragment which causes tissue-specific expression
in leaves, stems, cotyledons, and vascular tissue of the hypocotyl while causing
detectable levels of expression in root vascular tissue when operably
coupled directly to a DNA segment corresponding to the -72 to +8
promoter fragment of the Cauliflower Mosaic Virus 35S gene
- Use of the promoter as generally described above
- Plants cells and plants containing a chimeric construct in which the
promoter as generally described above is operably coupled to a
structural gene.
|
Claim 1
In a method for the expression of a chimeric plant gene, the improvement which
comprises the use of a tissue-specific promoter fragment which causes
tissue-specific expression in leaves, stems, cotyledons, and vascular tissue of
the hypocotyl while causing detectable levels of expression in root vascular
tissue when operably coupled directly to a DNA segment corresponding to the -72
to +8 promoter fragment of the Cauliflower Mosaic Virus 35S gene, said
tissue-specific promoter fragment having the sequence:
5'-TGAGACTTTT CAACAAAGGG TAATATCCGG AAACCTCCTC GGATTCCATT GCCCAGCTAT
CTGTCACTTT
ATTGTGAAGA TAGTGGAAAA GGAAGGTGGC TCCTACAAAT GCCATCATTG CGATAAAGGA AAGGCCATCG
TTGAAGATGC CTCTGCCGAC AGTGGTCCCA AAGATGGACC CCCACCCCAC GAGGAGCATC GTGGAAAAAG
AAGACGTTCC AACCACGTCT TCAAAGCAAG TGGATTGATG TGATA-3'*
*sequence corresponds to the complete domain B -from -343 to -90 nucleotides-
of the 35S CaMV promoter
|
|
Claim 2
A chimeric plant gene comprising in sequence in the 5' to 3' direction a
tissue-specific promoter fragment consisting essentially of the sequence:
5'-TGAGACTTTT CAACAAAGGG TAATATCCGG AAACCTCCTC GGATTCCATT GCCCAGCTAT
CTGTCACTTT
ATTGTGAAGA TAGTGGAAAA GGAAGGTGGC TCCTACAAAT GCCATCATTG CGATAAAGGA AAGGCCATCG
TTGAAGATGC CTCTGCCGAC AGTGGTCCCA AAGATGGACC CCCACCCCAC GAGGAGCATC GTGGAAAAAG
AAGACGTTCC AACCACGTCT TCAAAGCAAG TGGATTGATG TGATA-3'*,
operably coupled directly to the -72 to +8 promoter fragment of the CaMV 35S
gene, said -72 to +8 promoter fragment operably coupled to a structural gene.
*sequence corresponds to the complete domain B from -343 to -90 nucleotides
of the 35S CaMV promoter.
|
|
Claim 5
A tissue-specific promoter fragment which functions in plants to cause
tissue-specific expression in the leaves, stems, cotyledons and the vascular
tissue of the hypocotyl and detachable levels of expression in root vascular
tissue operably coupled directly to a DNA segment corresponding to the -72 to +8
promoter fragment of the Cauliflower Mosaic Virus 35S gene, said tissue-specific
promoter fragment having the sequence from its 5' to 3' termini:
5'-TGAGACTTTT CAACAAAGGG TAATATCCGG AAACCTCCTC GGATTCCATT GCCCAGCTAT
CTGTCACTTT
ATTGTGAAGA TAGTGGAAAA GGAAGGTGGC TCCTACAAAT GCCATCATTG CGATAAAGGA AAGGCCATCG
TTGAAGATGC CTCTGCCGAC AGTGGTCCCA AAGATGGACC CCCACCCCAC GAGGAGCATC GTGGAAAAAG
AAGACGTTCC AACCACGTCT TCAAAGCAAG TGGATTGATG TGATA-3'*
*sequence corresponds to the complete domain B -from -343 to -90 nucleotides-
of the 35S CaMV promoter.
|
|
Note: Patent information on this page was last updated on 15 March 2006.
Enforcement of rights
The 35S promoter is one of the most widely used promoters for driving the
expression of genes in transgenic plants.
Entities outside the U.S. and Japan using the promoter even
without a permit or license from the patent owners do
not infringe the patents, because patents are jurisdiction-specific.
However, laboratories in public and private institutes around the world using
the promoter as claimed in these patents in their experiments and in products
released into the market should bear in mind that unauthorized use or
import into the U.S. or Japan could constitute infringement.
While the patent owners have not shown interest in prosecuting academic users
of the promoter, they could choose to seek injunctions, damages and royalties
for infringing uses, including use in any commercial product. For instance,
Monsanto Company sued DNA Plant Technology Corporation in the U.S. for
infringing the U.S patent
5352605
by using the CaMV promoter in making and commercializing hybrid tomatoes with
delayed ripening. Before going out of business, DNA Plant Technology
Corporation had to acknowledge the infringement of this patent and also other
Monsanto patents, and entered into a confidential legal settlement agreement
that provided Monsanto with compensation.
Enhancer of CaMV 35S promoter
Scientific aspects
The subdomain B of the 35S promoter harbors an enhancer element that
increases promoter activity. Initially, enhanced transcription in transgenic
tobacco plants was obtained by duplicating the region from the -343 position to
the -90 position, which is upstream of the TATA sequence. The sequences involved
in the enhancement of transcription were localized to a 162 bp sequence
fragment, from -208 to -46 bp. Like other enhancers, this fragment can function
in an orientation-independent manner when located either upstream or downstream
of a homologous or heterologous TATA box. The region contains binding sites for
cellular trans-acting factors such as ASF 1, GATA 1 and CAF. An increase in
transcription activity from three-fold up to six-fold is reported in monocot
plants transformed with multiple copies of the enhancer region.
IP issues
The University of British Columbia was the initial applicant for several
patents covering the use of a duplicated CaMV 35S enhancer sequence as part of a
DNA construct for plant transformation. Monsanto Company is now
the assignee of all the patents granted in the U.S. and in Canada . The assignee
of the granted Brazilian patent is not identified, according to the information
provided by the International Patent Documentation Center (INPADOC). If you are
aware of updated licensing information, please provide it to other users via our
comments interface.
How is enhancer defined?
The applicant defines an enhancer as a sequence which activates transcription
and contains repetitive units of short sequences from about 4 to 16 bp. A
repetitive unit includes a GTGG, where G may be replaced by A in some cases. A
"natural enhancer" is described as comprising a DNA sequence that is upstream
from and within about 600 bp of a promoter.
Nowadays, it is known that enhancers can
be located thousands of base pairs upstream or downstream from the gene they
control. They can even be located within the gene under control. In the
specification, a promoter is described as the transcription initiation domain,
that is, a sequence that contains the TATA box and the CAP site. So, a
transcription initiation region is formed by a transcription initiation domain
or promoter and enhancer domain. In this way, the inventors divide a
"full-length" or " complete" 35S promoter into two main areas, and the enhancer
region becomes part of what is regarded in general as the promoter region.
Approximate scope of protection
In all the patents, the claims generally cover a fragment of the 35S upstream
region, defined as AluI-EcoRV fragment. This is described as a
truncated 35S enhancer, from -287 to -90 position. In the U.S., the claims are
directed to:
- A DNA construct having:
- a duplicated 35S enhancer, which corresponds to a duplication of the
truncated 35S enhancer;
- a promoter (with TATA box and CAP site);
- a sequence of interest; and
- a termination region.
- A plant cell having the mentioned construct.
- A chimeric transcriptional initiation region that, by having a tandemly
duplicated 35S enhancer, "enhances" the amount of transcription compared to
having one copy of the enhancer.
- A differentiated plant having the described chimeric transcriptional
initiation region.
- A method for expressing a gene of interest by transforming a plant cell with
the described chimeric transcriptional initiation region.
The Canadian patent claims the same type of DNA construct as
the U.S. patents. Also claimed are:
- a chimeric transcriptional initiation region contains two to four copies of
the truncated 35S enhancer;
- a method for producing a plant cell with enhanced expression of a gene of
interest using the multicopy truncated 35S enhancer; and
- a plant cell having a DNA construct with two copies of the truncated 35S
enhancer.
In addition, there are broader claims that encompass a transcription
initiation region with a plant or viral enhancer domain having four units of
enhancer sequences and a promoter. A DNA construct comprising such a
transcription initiation region is also claimed. The patent numbers and a
summary of the independent claims of each patent are presented
in the following table.
Patents on duplicated CaMV 35S enhancer sequences
|
Patent number
|
Title, Independent Claims and Summary of Claims
|
Assignee
|
|
US
5164316
- Earliest priority - 13 January 1987
- Filed - 17 August 1989
- Granted - 17 November 1992
- Expected expiry - 17 November 2009
|
Title - DNA construct for enhancing the efficiency of
transcription
| Claim 1
A plant cell comprising: a DNA construct having as components, (a) a
duplicated CaMV 35s enhancer sequence comprising an AluI-EcoRV
fragment of a CaMV 35S upstream region; and (ii) a promoter comprising an RNA
polymerase binding site and an mRNA initiation site; (b) a nucleotide sequence
of interest for transcription to mRNA; and (c) a termination region wherein said
components are operably joined.
|
|
Listed in the patent documents as the University of
British Columbia, but assigned to Monsanto in 1993
|
|
US
5196525
- Earliest priority - 13 January 1987
- Filed - 8 April 1991
- Granted - 23 March 1993
- Expected expiry -23 March 2010
|
Title - DNA construct for enhancing the efficiency of
transcription
Claim 1
A DNA construct having as components, (a) a transcription
initiation region including (i) a tandemly duplicated CaMV 35S
enhancer sequence comprising an AluI-EcoRV fragment of a CaMV 35S
upstream region; (ii) a promoter comprising an RNA polymerase binding site and
an mRNA initiation site; (b) a nucleotide sequence of interest for transcription
to mRNA; and (c) a termination region wherein said components are operably
joined.
|
|
|
US
5322938
- Earliest priority - 13 January 1987
- Filed - 17 November 1992
- Granted - 21 June 1994
- Expected expiry - 21 June 2011
|
Title - DNA sequence for enhancing the efficiency of
transcription
Claim 1
A chimeric transcriptional initiation region comprising:
as operably joined components (i) a tandemly
duplicated CaMV 35S enhancer sequence comprising an AluI-EcoRV
fragment of a CaMV 35S upstream region; and (ii) a promoter comprising an RNA
polymerase binding site and an mRNA initiation site, wherein when a nucleotide
sequence of interest is transcribed under the regulatory control of said
chimeric transcriptional initiation region, the amount of transcription product
is enhanced as compared to the amount of transcription obtained
with a chimeric transcriptional initiation region comprising a single copy of
said CaMV enhancer sequence and said promoter.
|
|
Monsanto
|
|
US
5359142
- Earliest priority - 13 January 1987
- Filed - 9 March 1994
- Granted - 25 October 1994
- Expected expiry - 25 October 2011
|
Title - Method for enhanced expression of a protein
Claim 1
A differentiated plant comprising that comprise a DNA construct having as
components (a) a tandemly duplicated CaMV 35S enhancer sequence comprising an
AluI-EcoRV fragment of a CaMV 35S upstream region; and (b) a promoter
comprising an RNA polymerase binding site and an mRNA initiation site; (c) a
nucleotide sequence of interest; and (d) a termination region; wherein said
components are operably joined.
|
This patent is a divisional of
US
5322938.
- A differentiated plant comprising the DNA construct as generally claimed in
US
5196525.
|
|
US
5424200
- Earliest priority - 13 January 1987
- Filed - 11 July 1994
- Granted - 13 June 1995
- Expected expiry - 13 June 2012
|
Title - Method for enhanced expression of a DNA sequence of
interest
|
Claim 1
A method for expressing a DNA sequence of interest comprising
transforming a plant cell with a DNA construct comprising (a) a tandemly
duplicated CaMV 35S enhancer sequence comprising an AluI-EcoRV
fragment of a CaMV 35S upstream region; (b) a promoter comprising an RNA
polymerase binding site and an mRNA initiation site; (c) a nucleotide sequence
of interest; and (d) a termination region; wherein said components are operably
joined so that said sequence of interest is transcribed.
|
This patent is a divisional of
US
5359142.
- A method for expressing a DNA sequence of interest by transforming a plant
cell with a DNA construct as claimed in
US
5196525.
|
|
CA 1325191
- Earliest priority -- 13 January 1987
- Filed - 12 January 1988
- Granted - 14 December 1993
- Expected expiry - 14 December 2010
|
Title - DNA construct for enhancing the efficiency of
transcription
Claim 1
A transcriptional initiation region comprising:
(a) a plant or viral
enhancer domain comprising a plurality of units each including a sequence of the
group comprising: GTGG, ATGG, GTAG, GTGA, GTGGA(T)A(T)A(T),GTGTGGA(T)A(T)A(T)G,
and complementary sequence thereof, said enhancer domain further comprising at
least one additional unit including a sequence selected from the said group; and
(b) a transcription initiation domain comprising an RNA polymerase binding
site and an mRNA initiation site, under the enhancing control of said enhancer
domain.
|
Claim 6
A DNA construct comprising: a transcription initiation region comprising:
(a) an enhancer domain comprising a plurality of units each including a
sequence selected from the group comprising: GTGG, ATGG, GTAG, GTGA,
GTGGA(T)A(T)A(T),GTGTGGA(T)A(T)A(T)G, and complementary sequence thereof; and
(b) a transcription initiation domain comprising an RNA polymerase binding
site and an mRNA initiation site under the enhancing control of said enhancer
domain;
(c) a sequence of interest for transcription to mRNA; and
(d) a termination region.
|
Claim 22
A DNA sequence comprising a transcription initiation region capable of
regulating transcription of a nucleotide sequence of interest in a plant cell
and including, as operably joined components, (i) two to four copies in tandem
of an enhancer sequence comprising an AluI-EcoRV fragment of a CaMV 35S
upstream region; and (ii) a promoter comprising an RNA polymerase binding site
and an mRNA initiation site.
|
Claim 29
A chimeric transcriptional initiation region comprising: as operably joined
components, (i) two or more copies of a CaMV 35S enhancer sequence and (ii) a
promoter comprising an RNA polymerase binding site and an mRNA initiation site,
wherein when a nucleotide sequence of interest is transcribed under the
regulatory control of said chimeric transcriptional initiation region, the
amount of transcription product is enhanced as compared to the amount of
transcription obtained with a chimeric transcriptional initiation region
comprising a single copy of said CaMV enhancer sequence and a promoter.
|
Claim 33
A DNA construct having as components, (a) a transcription initiation region
including (i) a tandemly duplicated CaMV 35S enhancer sequence comprising an
AluI-EcoRV fragment of a CaMV 35S upstream region; (ii) a promoter comprising an
RNA polymerase binding site and an mRNA initiation site; (b) a nucleotide
sequence of interest for transcription to mRNA; and (c) a termination region
wherein said components are operably joined.
|
Claim 38
A method for producing a plant cell capable of enhanced
expression of a nucleotide sequence of interest, said method
comprising: operably linking to form an expression cassette
said nucleotide sequence of interest and a transcription initiation region
capable of regulating transcription of a DNA sequence of interest in a plant
cell, wherein said transcription initiation region comprises (i) two to four
copies in tandem of an enhancer sequence comprising an AluI-EcoRV
fragment of a CaMV 35S upstream region; and (ii) a promoter comprising an RNA
polymerase binding site and an mRNA initiation site; and introducing said
expresssion cassette into said plant cell to form a plant cell capable of
enhanced expression of a said nucleotide sequence of interest.
|
Claim 47
A plant cell comprising: a DNA construct having components, (a) a
transcription initiation region including (i) a tandemly duplicated CaMV 35S
enhancer sequence comprising an AluI-EcoRV fragment of a CaMV 35S
upstream region; (ii) a promoter comprising an RNA polymerase binding site and
an mRNA initiation site; (b) a nucleotide sequence of interest for transcription
to mRNA; and (c) a termination region wherein said components are operably
joined.
|
Claim elements:
- A DNA transcription region comprising 2 to 4 copies in tandem of enhancer
35S and a promoter region as generally claimed in
US
5164316.
- A chimeric transcriptional initiation region comprising 2 or more copies of
enhancer 35S and having the properties as generally claimed in
US
5322938.
- A DNA construct as generally claimed in
US
5196525.
- A method of producing a plant cell capable of enhanced
expression of a gene of interest by having in an expression cassette with a gene
of interest linked to a DNA transcription region as described in the first
point.
- A plant cell comprising a DNA construct as generally claimed in
US
5164316.
|
|
Remarks
|
A related patent has been granted in Brazil (BR 1101045).
|
|
Note: Patent information on this page was last updated on 6 March 2006.
Challenges to patents on 35S promoter and duplicated CaMV 35S enhancer
sequences
In September 2006, PUBPAT filed formal
requests with the United States Patent and Trademark Office to revoke four
patents owned by Monsanto Company. PUBPAT has issued a
press release giving
the opinion that these patents were a subject for action in the public interest
because Monsanto is using these patents to intimidate and sue farmers.
The patents are U.S. Patents
5352605,
5164316,
5196525,
and
5322938,
all of which assert claims to the CaMV 35S promoter, double 35S enhancer, and/or
constructs containing them. Like some other Monsanto technologies, the 35S
promoter and double 35S enhancer have been used in a great deal of agricultural
research worldwide, and as a result Monsanto's intellectual property does
constrain the delivery of products.
In its filings, PUBPAT submitted putative prior art supporting an assertion
that the patents should not have been granted, and therefore should be revoked.
The USPTO will do a re-examination of the patents in view of the submission, and
if it is found that the patents are invalid over the prior art, may require
revision or revocation of specified claims. The independent claims are listed
below for easy reference.
|
Patent Number
|
Title, Independent Claims and Summary of Claims
|
Assignee
|
|
US
5352605
- Earliest priority - 17 January 1983
- Filed - 28 October 1993
- Granted - 4 October 1994
- Expected expiry - 4 October 2011
|
Title - Chimeric genes for transforming plant cells using
viral promoters
Claim 1
A chimeric gene which is expressed in plant cells comprising a promoter from
a cauliflower mosaic virus, said promoter selected from the group consisting of
a CaMV (35S) promoter isolated from CaMV protein-encoding DNA sequences and a
CaMV (19S) promoter isolated from CaMV protein-encoding DNA sequences, and a
structural sequence which is heterologous with respect to the promoter.
|
Claim 4
A plant cell which comprises a chimeric gene that contains a promoter from
cauliflower mosaic virus, said promoter selected from the group consisting of a
CaMV (35S) promoter and a CaMV (19S) promoter, wherein said promoter is isolated
from CaMV protein-encoding DNA sequences, and a structural sequence which is
heterologous with respect to the promoter.
|
Claim 7
An intermediate plant transformation plasmid which comprises a region of
homology to an Agrobacterium tumefaciens vector, a T-DNA border region
from Agrobacterium tumefaciens and a chimeric gene, wherein the
chimeric gene is located between the T-DNA border and the region of homology,
said chimeric gene comprising a promoter from cauliflower mosaic virus, said
promoter selected from the group consisting of a CaMV(35S) promoter and a
CaMV(19S) promoter, and a structural sequence which is heterologous with respect
to the promoter.
|
Claim 8
A plant transformation vector which comprises a disarmed plant tumor inducing
plasmid of Agrobacterium tumefaciens and a chimeric gene, wherein the
chimeric gene contains a promoter from cauliflower mosaic virus, said promoter
selected from the group consisting of a CaMV(35S) promoter and a CaMV(19S)
promoter, and a structural sequence which is heterologous with respect to the
promoter.
|
Claim 13
A DNA construct comprising: (A) a CaMV promoter selected from the group
consisting of (1) a CaMV 35S promoter isolated from CaMV protein-encoding DNA
sequences and (2) a CaMV 19S promoter isolated from CaMV protein-encoding DNA
sequences, and (B) a DNA sequence of interest heterologous to (A), wherein (B)
is under the regulatory control of (A) when said construct is transcribed in a
plant cell.
|
Claim 14
A chimeric gene which is transcribed and translated in plant cells, said
chimeric gene comprising a promoter from cauliflower mosaic virus, said promoter
selected from the group consisting of: a) a CaMV 35S promoter region free of
CaMV protein-encoding DNA sequences and b) a CaMV 19S promoter region free of
CaMV protein-encoding DNA sequences, and a DNA sequence which is heterologous
with respect to the promoter.
|
Claim 15
A chimeric gene which is expressed in plants cells comprising a promoter from
a cauliflower mosaic virus, said promoter selected from the group consisting of
a CaMV(35S) promoter region free of CaMV protein-encoding DNA sequences and a
CaMV(19S) promoter region free of CaMV protein-encoding DNA sequences, and a DNA
sequence which is heterologous with respect to the promoter.
|
Claim 16
A chimeric gene which is transcribed in plants cells comprising a promoter
from a cauliflower mosaic virus, said promoter selected from the group
consisting of a CaMV(35S) promoter free of CaMV protein-encoding DNA sequences
and a CaMV(19S) promoter free of CaMV protein-encoding DNA sequences, a DNA
sequence which is heterologous with respect to the promoter and a 3'
non-translated polyadenylation signal sequence.
|
Claim 17
A plant cell which comprises a chimeric gene where said chimeric gene
comprises a promoter from cauliflower mosaic virus, said promoter selected from
the group consisting of a CaMV(35S) promoter and a CaMV(19S) promoter, wherein
said promoter is free of CaMV protein-encoding DNA sequences, and a DNA sequence
which is heterologous with respect to the promoter and a 3' non-translated
polyadenylation signal sequence.
|
Claim elements:
- 35S and 19S promoters, as defined in the specification, are
claimed as part of a chimeric construct, also containing a heterologous signal
and a poly(A) signal, expressed in a plant
- There are also claims to intermediate plant transformation vectors and plant
transformation vectors containing the chimeric gene for
Agrobacterium-mediated plant transformation, and to plant cells having
the chimeric gene.
|
Monsanto
|
|
US
5164316
- Earliest priority - 13 January 1987
- Filed - 17 August 1989
- Granted - 17 November 1992
- Expected expiry - 17 November 2009
|
Title - DNA construct for enhancing the efficiency of
transcription
|
Claim 1
A plant cell comprising a DNA construct having as
components,
(a) a duplicated CaMV 35s enhancer sequence comprising
an AluI-EcoRV fragment of a CaMV 35S upstream region;
and
(ii) a promoter comprising an RNA polymerase binding site and an mRNA
initiation site;
(b) a nucleotide sequence of interest for transcription to mRNA;
and
(c) a termination region wherein said components are operably
joined.
|
|
Listed in the patent documents as the University of
British Columbia, but assigned to Monsanto in 1993
|
|
US
5196525
- Earliest priority - 13 January 1987
- Filed - 8 April 1991
- Granted - 23 March 1993
- Expected expiry -23 March 2010
|
Title - DNA construct for enhancing the efficiency of
transcription
|
Claim 1
A DNA construct having as components,
(a) a transcription initiation region including
(i) a tandemly duplicated CaMV 35S enhancer sequence
comprising an AluI-EcoRV fragment of a CaMV 35S
upstream region;
(ii) a promoter comprising an RNA polymerase binding site and an mRNA
initiation site;
(b) a nucleotide sequence of interest for transcription to mRNA;
and
(c) a termination region wherein said components are operably
joined.
|
|
|
US
5322938
- Earliest priority - 13 January 1987
- Filed - 17 November 1992
- Granted - 21 June 1994
- Expected expiry - 21 June 2011
|
Title - DNA sequence for enhancing the efficiency of
transcription
|
Claim 1
A chimeric transcriptional initiation region comprising:
as operably joined components
(i) a tandemly duplicated CaMV 35S enhancer sequence
comprising an AluI-EcoRV fragment of a CaMV 35S upstream region;
and
(ii) a promoter comprising an RNA polymerase binding site and an mRNA
initiation site, wherein when a nucleotide sequence of interest is transcribed
under the regulatory control of said chimeric transcriptional initiation region,
the amount of transcription product is enhanced as compared to
the amount of transcription obtained with a chimeric transcriptional
initiation region comprising a single copy of said CaMV enhancer sequence and
said promoter.
|
|
|
Remarks
|
Corresponding applications also be pending and there may be issued claims in
Brazil and Japan.
|
Note: Patent information on this page was last updated on 30 September 2006.
CaMV 35S minimal promoter
Scientific aspects
Part of the domain A of the CaMV 35S promoter, which contains the TATA box
and extends from the -90 position to the transcription start site +1, is used by
many as a "minimal promoter." Apart from the TATA box, which is
the binding site for RNA polymerase II, the region contains a least three
CAAT-like boxes. These sequences potentiate the activity of upstream sequences
and influence the efficiency of the promoter activity. These CAAT-like boxes
alone, or attached to heterologous promoter regions, drive the expression of
transgenes.
The so-called minimal promoter, which is often described as a sequence
similar to that extending from nucleotides -60 or -46 to +1 of the 35S promoter,
does not appear to drive the expression of a gene by itself. Additional
sequences, such as an enhancer, are required. Thus, the 35S minimal promoter
has often been used to find and define enhancer regions, and determine activity
of heterologous promoters, and as part of trans-activating or inducible
promoter systems.
IP issues
A careful reading of the patents on the CaMV 35S promoter, including the ones
directed to the subdomains of the promoter and the enhancer region, suggests but
does not confirm that the widely used "minimal promoter" may be unencumbered by
patents. The most relevant claims to the 35S minimal promoter were granted to
the Rockefeller University in the patent
US
5097025. Several independent claims cover the minimal promoter of
the CaMV 35S promoter, which the inventors define as sequence from -46 to +8,
but the minimal promoter is not claimed by itself, rather as linked to
one of the subdomains of domain B of the 35S promoter
Because of the minimal amount of sequence actually derived from the 35S
promoter, we suggest that scientists refer to it simply as a "minimal promoter."
Opine Promoters
Scientific Aspects
Under natural conditions, opine promoters drive the expression of opines
(i.e. mannopine, octopine, nopaline), hormone-like compounds generated by the
soil bacterium Agrobacterium through the use of the plant's expression
machinery. Opines are utilized by the bacterium as a source of carbon, nitrogen
and energy. Promoters from the nopaline synthase (nos), octopine
synthase (ocs) and mannopine synthase (mas) genes have been
isolated and inserted into transformation vectors upstream of foreign genes
to control the expression of those genes.
Although these promoters are regarded as constitutive, their level of
activity can be affected by hormones and wounding. Nevertheless, they are still
frequently used, mainly for transformation of dicotyledonous (dicot) plants.
Additionally, depending on the orientation, some functional parts of regulatory
sequences associated with promoters, such as the mas promoters, have
the ability to bind nuclear protein factors from different plants and can be
used as enhancers or silencers.
IP issues
Granted patents related to transcriptional regulatory elements derived from
opine synthase genes may be divided into three groups:
-
Promoters and UAS from opine synthase genes: Patents
granted in the U.S. and Australia directed to promoters and upstream activating
sequences (UAS) derived from opine synthase genes, especially from the
ocs and mas genes of A. tumefaciens;
-
35S-enhanced mas promoter: Patents granted in the
U.S. and Canada directed to a mas promoter enhanced by a UAS from CaMV
35S gene; and
-
Enhancers from the ocs gene: Patents granted in the
U.S., Europe and Canada directed to enhancer sequences isolated from the
ocs gene of particular use in the transformation of monocots.
Promoters and UAS from opine synthase genes
The Biotechnology Research and Development Corporation and the Purdue
Research Foundation co-own patents granted in the U.S., Australia and Russia.
The United States and the Australian patents are directed to chimeric regulatory
regions containing promoters and UAS derived from the opine genes mas
and ocs of Agrobacterium tumefaciens. The UAS are essential for the
transcription of the opine genes in natural conditions. The promoters of
mas and ocs are themselves weak promoters, but linking UAS
regions to them increases their activity and they behave in a more constitutive
fashion.
Definition of promoter and UAS
A UAS is defined in these patents in functional terms as a
cis-acting element that in the native state is usually at least 100 bp
upstream of the native transcriptional start site and can exert influence on
gene expression.
A promoter is not precisely defined in the specification, but it is referred
to as a DNA sequence required for the transcription of genes that contains a
functional TATA box and which directs the expression of a gene. Thus, the
promoter has both structural and functional elements.
Approximate scope of protection
The chimeric regulatory regions claimed in the U.S. patent combine promoters
and UAS elements as follows:
- a promoter and an UAS from a mas gene and at least one UAS from an
ocs gene;
- a promoter from a mas gene and at least 3 UAS elements from an
ocs gene; and
- a promoter and an UAS from a mas gene and at least 3 UAS
elements from an ocs gene.
The Australian patent claims more combinations of promoters and UAS elements
than the U.S. patent. In contrast with the U.S. patent, the promoters and UAS of
some of the regulatory regions claimed in the Australian patent derive from
any opine synthase gene. It also includes a claim to a
regulatory region formed by a promoter and a UAS, both derived from a
mas gene.
Cassettes and methods for the expression of a foreign gene in a plant using
the described chimeric regulatory regions are also claimed in both the U.S. and
the Australian patents. Some of the cassettes are used for
inducible expression of a gene of interest in a plant.
Transgenic plants containing some of the cassettes are also claimed.
Some bibliographic data and a summary of the independent
claims of the granted patents are presented in the table on the following page.
Promoters and UAS from opine synthase genes - Specific Patent Information
|
Patent Number
|
Title, Independent Claims and Summary of
Claims
|
Assignee
|
|
US
5955646
- Earliest priority - 19 November 1993
- Filed - 31 July 1996
- Granted - 21 September 1999
- Expected expiry - 19 November 2013
|
Title - Chimeric regulatory regions and gene cassettes for
expression of genes in plants
| Claim 1
A cassette for inducible expression of a foreign gene
comprising said foreign gene operably linked to a regulatory
region comprising
a promoter derived from a mannopine
synthase gene of Agrobacterium tumefaciens,
an upstream
activating sequence derived from a mannopine synthase gene of Agrobacterium
tumefaciens, and
at least one upstream activating sequence
derived from an octopine synthase gene of Agrobacterium tumefaciens.
|
| Claim 2
A method for nematode inducible expression of a foreign gene in a plant,
comprising:
linking said foreign gene to a regulatory region
comprising a promoter derived from a mannopine synthase gene of
Agrobacterium tumefaciens comprising 138 bases
upstream of the transcription initiation site, and an upstream activating
sequence derived from a mannopine synthase gene of Agrobacterium
tumefaciens;
inserting said foreign gene and said regulatory
region in said plant, wherein expression is induced by nematode attack on the
plant.
|
Claim 3
A method for nemotode inducible expression of a foreign gene in a plant,
comprising:
linking said foreign gene to a
regulatory region comprising
a promoter derived
from a mannopine synthase gene of Agrobacterium tumefaciens,
an upstream activating sequence derived from a mannopine synthase gene of
Agrobacterium tumefaciens, and
at least one upstream
activating sequence derived from an octopine synthase gene of Agrobacterium
tumefaciens;
inserting said foreign gene and said regulatory
region in said plant, wherein expression is induced by nematode attack on the
plant.
|
Claim 4
A chimeric regulatory region for expressing genes in plants
comprising at least three upstream activating sequences derived
from an Agrobacterium tumsfaciens octopine synthase gene operably
linked to a promoter derived from an Agrobacterium tumefaciens
mannopine synthase gene.
|
Claim 5
A cassette for expressing a foreign gene comprising the
foreign gene operably linked to a chimeric regulatory region
comprising at least three upstream activating sequences derived
from Agrobacterium tumefaciens octopine synthase genes operably linked
to a promoter derived from an Agrobacterium tumefaciens mannopine
synthase gene.
|
Claim 6
A plasmid comprising a cassette comprising
a foreign gene operably linked to a chimeric regulatory region
comprising at least three upstream activating sequences derived
from Agrobacterium tumefaciens octopine synthase genes operably linked
to a promoter derived from an Agrobacterium tumefaciens mannopine
synthase gene.
|
Claim 7
A method of expressing a foreign gene in a plant,
comprising:
linking said foreign gene to a chimeric
regulatory region comprising at least three upstream activating
sequences derived from an Agrobacterium tumefaciens octopine synthase
gene operably linked to a promoter derived from an Agrobacterium
tumefaciens mannopine synthase gene; and inserting said foreign
gene and said chimeric regulatory region into a plant, wherein said plant
expresses said foreign gene.
|
Claim 8
A transgenic plant comprising a cassette
comprising a foreign gene operably linked to a chimeric
regulatory region comprising at least three upstream activating
sequences derived from Agrobacterium tumefaciens octopine synthase
genes operably linked to a promoter derived from an Agrobacterium
tumefaciens mannopine synthase gene.
|
Claim 9
A chimeric regulatory region for expressing genes in plants
comprising at least three upstream activating sequences derived
from an Agrobacterium tumefaciens octopine synthase gene operably
linked to an upstream activating sequence derived from an Agrobacterium
tumefaciens mannopine synthase gene that is operably linked to a promoter
derived from an Agrobacterium tumefaciens mannopine synthase gene.
|
Claim 10
A cassette for expressing a foreign gene comprising the
foreign gene operably linked to a chimeric regulatory region
comprising at least three upstream activating sequences derived
from Agrobacterium tumefaciens octopine synthase genes operably linked
to an upstream activating sequence derived from an Agrobacterium
tumefaciens mannopine synthase gene that is operably linked to a promoter
derived from an Agrobacterium tumefaciens mannopine synthase gene.
|
Claim 11
A method of expressing a foreign gene in a plant,
comprising:
linking said foreign gene to a chimeric
regulatory region comprising at least three upstream activating
sequences derived from an Agrobacterium tumefaciens octopine synthase
gene operably linked to an upstream activating sequence derived from an
Agrobacterium tumefaciens mannopine synthase gene that is operably
linked to a promoter derived from an Agrobacterium tumefaciens
mannopine synthase gene; and
inserting said foreign gene and said
chimeric regulatory region into a plant, wherein said plant expresses said
foreign gene.
|
Claim 12
A transgenic plant comprising a chimeric regulatory region
for expressing genes in plants comprising at least three
upstream activating sequences derived from an Agrobacterium tumefaciens
octopine synthase gene operably linked to an upstream activating sequence
derived from an Agrobacterium tumefaciens mannopine synthase gene that
is operably linked to a promoter derived from an Agrobacterium
tumefaciens mannopine synthase gene.
|
The claims are directed to:
- A cassette for inducible expression of a foreign gene operably linked to a
regulatory region having
- a promoter and an UAS from the mas gene and
- at least one UAS from an ocs gene.
The term "operably linked" is not explicitly defined but in the
context of the disclosure appears to refer to a gene sequence present on the
same nucleic acid molecule as a promoter sequence and whose expression is under
the control of that promoter sequence.
- A chimeric regulatory region for expressing genes in plants having at least
3 UAS derived from an ocs gene linked to a promoter derived from a
mas gene. A cassette, a plasmid, a method of expressing a foreign gene
in a plant using this regulatory region, methods for nematode inducible
expression of a foreign gene, and a transgenic plant are also claimed.
- A chimeric regulatory region for expressing genes in plants having
- 4 UASs,
- at least 3 UASs from an ocs gene and one UAS from a mas gene linked
to a mas gene promoter.
- A cassette, a method of expressing a foreign gene in a plant using this
regulatory region and a transgenic plant are also claimed.
The disclosure describes that a "foreign gene" includes "any DNA
that is sought to be expressed in [a] transgenic plant. In this context, the
gene, no matter the source, is inserted into the plant genome and is thus
foreign to that plant in the location of insertion, even if the gene originated
from the plant being transformed." So a foreign gene can be a plant gene and may
even be from the plant being transformed if inserted into a different genomic
location from where it is usually found.
A cassette, as described in the patent, does not necessarily include an
origin of replication. As claimed, a cassette could encompass a plasmid or a
portion of a plasmid (e.g., a linear fragment).
|
Biotechnology
Research and Development Corporation
& Purdue
Research Foundation
|
|
AU 687961
B
- Earliest priority - 19 November 1993
- Filed - 17 November 1994
- Granted - 5 March 1998
- Expected expiry - 16 November 2014
|
Title - Chimeric regulatory regions and gene cassettes for
expression of genes in plants
| Claim 1
A chimeric regulatory region for expressing genes in plants
comprising an upstream activating sequences derived from an
Agrobacterium tumefaciens octopine synthase gene operably linked to a
promoter derived from an Agrobacterium tumefaciens mannopine synthase
gene.
|
| Claim 2
A cassette for expressing a gene comprising a gene operably
linked to a chimeric regulatory region comprising an upstream
activating sequences derived from an Agrobacterium tumefaciens
octopine synthase gene operably linked to a promoter derived from an
Agrobacterium tumefaciens mannopine synthase gene.
|
| Claim 4
A chimeric regulatory region for expressing genes in plants
comprising at least two upstream activating sequences derived
from an Agrobacterium tumefaciens opine synthase gene operably linked
to a promoter derived from an Agrobacterium tumefaciens opine synthase
gene, wherein at least one of said upstream activating elements are derived from
a different opine synthase gene than said promoter.
|
| Claim 8
A cassette for expressing a gene comprising a gene operably
linked to a chimeric regulatory region comprising at least two
upstream activating sequences derived from an Agrobacterium tumefaciens
opine synthase gene operably linked to a promoter derived from an
Agrobacterium tumefaciens opine synthase gene, wherein at least one of
said upstream activating elements are derived from a different opine synthase
gene than said promoter.
|
| Claim 13
A cassette for inducible expression of a foreign gene
comprising said foreign gene operably linked to a regulatory
region comprising:
a) a promoter derived from a mannopine synthase gene of
Agrobacterium tumefaciens by deletion to nucleotide position -138, and
b) an upstream activating sequence derived from a mannopine synthase
gene of Agrobacterium tumefaciens.
|
| Claim 15
A cassette for inducible expression of a foreign gene
comprising said foreign gene operably linked to a regulatory
region comprising:
a) a promoter derived from a mannopine synthase gene of
Agrobacterium tumefaciens,
b) an upstream activating sequence
derived from a mannopine synthase gene of Agrobacterium tumefaciens,
and
c) an upstream activating sequence derived from an octopine
synthase gene of Agrobacterium tumefaciens.
|
| Claim 16
A method for expressing a gene in a plant, comprising the
steps of:
a) linking said gene to a chimeric regulatory region comprising an
upstream activating sequence derived from an Agrobacterium tumefaciens
octopine synthase gene operably linked to a promoter derived from an
Agrobacterium tumefaciens mannopine synthase gene;
b)
inserting said gene and said chimeric regulatory region into a plant; and
c) allowing said plant to express said gene.
|
|
Claim 17
A method for expressing a gene in a plant,
comprising the steps of:
a) linking said gene to a chimeric regulatory region comprising at least
two upstream activating sequences derived from an Agrobacterium tumefaciens
opine synthase gene operably linked to a promoter derived from an
Agrobacterium tumefaciens opine synthase gene, wherein at least one of
said upstream activating elements are derived from a different opine synthase
gene than said promoter;
b) inserting said gene and said chimeric
regulatory region into a plant; and
c) allowing said plant to express
said gene.
|
| Claim 21
A method of inducible expression of a foreign gene in a plant,
comprising:
a) linking said foreign gene to a regulatory region
comprising:
i. a promoter derived from a
mannopine synthase gene of Agrobacterium tumefaciens by deletion to
nucleotide position -138, and
ii. an upstream activating sequence
derived from a mannopine synthase gene of Agrobacterium tumefaciens;
b) inserting said foreign gene and said regulatory region in said
plant; and
c) inducing expression of said foreign gene.
|
| Claim 23
A method for inducible expression of a foreign gene in a plant,
comprising:
a) linking said foreign gene to a regulatory region
comprising:
i. a promoter derived from a mannopine
synthase gene of Agrobacterium tumefaciens,
ii. an upstream
activating sequence derived from a mannopine synthase gene of Agrobacterium
tumefaciens; and
iii. an upstream activating sequence derived
from an octopine synthase gene of Agrobacterium tumefaciens;
b) inserting said foreign gene and said regulatory region in said plant; and
c) inducing expression of said foreign gene.
|
| Claim 32
A chimeric regulatory region for expressing genes in plants
comprising at least three upstream activating sequences derived
from an Agrobacterium tumefaciens octopine synthase gene operably
linked to a promoter derived from an Agrobacterium tumefaciens
mannopine synthase gene.
|
| Claim 33
A cassette for expressing a gene comprising a gene operably
linked to a chimeric regulatory region comprising at least
three upstream activating sequences derived from Agrobacterium
tumefaciens octopine synthase genes operably linked to a promoter derived
from an Agrobacterium tumefacie ns mannopine synthase gene.
|
| Claim 34
A method of expressing a gene in a plant, comprising the
steps of:
a) linking said gene to a chimeric regulatory region comprising at least
three upstream activating sequences derived from an Agrobacterium
tumefaciens octopine synthase gene operably linked to a promoter derived
from an Agrobacterium tumefaciens mannopine synthase;
b)
inserting said gene and said chimeric regulatory region into a plant; and
c) allowing said plant to express said gene.
|
| Claim 37
A chimeric regulatory region for expressing a gene in a plant
comprising at least three upstream activating sequences derived
from an Agrobacterium tumefaciens opine synthase gene operably linked
to a promoter derived from an Agrobacterium tumefaciens opine synthase
gene, wherein at least one of said upstream activating elements are derived from
a different opine synthase gene than said promoter.
|
| Claim 39
A cassette for expressing a gene in a plant comprising a
gene operably linked to a chimeric regulatory region comprising
at least three upstream activating sequences derived from Agrobacterium
tumefaciens opine synthase genes operably linked to a promoter derived from
an Agrobacterium tumefaciens opine synthase gene, wherein at least one
of said upstream activating elements are derived from a different opine synthase
gene than said promoter.
|
| Claim 40
A chimeric regulatory region for expressing a gene in a plant
comprising at least three upstream activating sequences derived
from an Agrobacterium tumefaciens opine synthase gene operably linked
to a promoter derived from an Agrobacterium tumefaciens opine synthase
gene.
|
| Claim 41
A chimeric regulatory region for expressing a gene in a plant
comprising at least two upstream activating sequences derived
from an Agrobacterium tumefaciens opine synthase gene operably linked
to a promoter derived from an Agrobacterium tumefaciens opine synthase
gene.
|
The claim elements are:
- A chimeric regulatory region for expressing genes in plants comprising a
promoter derived from a mas gene and an UAS derived from an
ocs gene.
- A chimeric regulatory region for expressing genes in plants having at least
2 UAS elements and a promoter from an opine synthase gene. One of the UAS
elements derives from a different opine gene than the promoter.
- A cassette for inducible expression of a foreign gene linked to a regulatory
region having a promoter and a UAS from a mas gene.
- A cassette for inducible expression of a foreign gene linked to a regulatory
region having a promoter and a UAS from a mas gene and a UAS from an
ocs gene.
- A chimeric regulatory region for expressing genes in plants having at least
3 UAS elements from an ocs gene linked to a promoter from a
mas gene.
- A chimeric regulatory region for expressing genes in plants having at least
3 UAS elements and a promoter from an opine synthase gene. One of the UAS is
from a different opine gene than the promoter.
- Chimeric regulatory regions comprising at least 2 and at least 3 UAS
elements from an opine synthase gene and a promoter also from an opine synthase
gene.
- Cassettes containing the mentioned chimeric regulatory regions and methods
for expressing a gene in a plant are also claimed.
|
|
EP
729514 B1
- Earliest priority - 19 November 1993
- Filed - 4 September 1996
- Granted - 12 August 2005
- Expected expiry - 3 September 2016
|
Title - Chimeric regulatory regions and gene cassettes for
expression of genes in plants
The number of claims in this granted European patent was reduced to nine with
only one independent claim as follows:
| Claim 1
A chimeric regulatory region for expressing genes in plants
comprising an upstream activating sequence of an Agrobacterium
tumefaciens opine synthase gene operably linked to a promoter derived from a
second Agrobacterium tumefaciens opine synthase gene operably linked to a
promoter of an Agrobacterium tumefaciens mannopine synthase.
|
|
|
|
CA
2174954
- Earliest priority - 19 November 1993
- Filed - 24 April 1996
- Granted - 15 March 2005
- Expected expiry - 23 April 2016
|
Title - Chimeric regulatory regions and gene cassettes for
expression of genes in plants
| Claim 1
A chimeric regulatory region for expressing genes in plants
comprising an upstream activating sequence derived from an
Agrobacterium tumefaciens octopine synthase gene operably linked to a
promoter derived from an Agrobacterium tumefaciens mannopine synthase
gene.
|
Claim 2
A chimeric regulatory region for expressing genes in plants
comprising at least two upstream activating sequences derived
from an Agrobacterium tumefaciens opine synthase gene operably linked
to a promoter derived from an Agrobacterium tumefaciens opine synthase
gene, wherein at least one of said upstream activating elements are derived from
a different opine synthase gene than said promoter.
|
Claim 3
A chimeric regulatory region for expressing genes in plants
comprising at least three upstream activating sequences derived
from an Agrobacterium tumefaciens octopine synthase gene operably
linked to a promoter derived from an Agrobacterium tumefaciens
mannopine synthase gene.
|
Claim 7
A cassette for expressing a gene in plants, said cassette
comprising a gene operably linked to chimeric regulatory region
comprising an upstream activating sequence derived from an
Agrobacterium tumefaciens octopine synthase gene operably linked to a
promoter derived from an Agrobacterium tumefaciens mannopine synthase
gene.
|
Claim 8
A cassette for expressing a gene in plants, said cassette
comprising a gene operably linked to chimeric regulatory region
comprising at least two upstream activating sequences derived
from an Agrobacterium tumefaciens opine synthase gene operably linked
to a promoter derived from an Agrobacterium tumefaciens opine synthase
gene, wherein at least one of said upstream activating elements are derived from
a different opine synthase gene than said promoter.
|
Claim 12
A cassette for inducible expression of a foreign gene in plants, said
cassette comprising said foreign gene operably linked to a
regulatory region comprising
a promoter derived from
a mannopine synthase gene of Agrobacterium tumefaciens by deletion to
base pair position -138 and
an upstream activating sequence derived
from a mannopine synthase gene of Agrobacterium tumefaciens.
|
Claim 13
A cassette for inducible expression of a foreign gene in plants, said
cassette comprising said foreign gene operably linked to a
regulatory region comprising
a promoter derived from
a mannopine synthase gene of Agrobacterium tumefaciens,
an
upstream activating sequence derived from a mannopine synthase gene of
Agrobacterium tumefaciens, and
an upstream activating
sequence derived from an octopine synthase gene of Agrobacterium
tumefaciens.
|
Claim 14
A cassette for expressing a gene in plants, said cassette
comprising a gene operably linked to a chimeric regulatory
region comprising at least three upstream activating sequences
derived from Agrobacterium tumefaciens octopine synthase genes operably
linked to a promoter derived from an Agrobacterium tumefaciens
mannopine synthase gene.
|
Claim 16
A method for expressing a gene in a plant, comprising the
steps of:
linking said gene to a chimeric regulatory region
comprising an upstream activating sequence derived from an
Agrobacterium tumefaciens octopine synthase gene operably linked to a
promoter derived from an Agrobacterium tumefaciens mannopine synthase
gene;
inserting said gene and said chimeric regulatory region into a
plant; and
allowing said plant to express said gene.
|
Claim 17
A method for expressing a gene in a plant, comprising the
steps of:
linking said gene to a chimeric regulatory region
comprising at least two upstream activating sequences derived
from an Agrobacterium tumefaciens opine synthase gene operably linked
to a promoter derived from an Agrobacterium tumefaciens opine synthase
gene, wherein at least one of said upstream activating elements are derived from
a different opine synthase gene than said promoter;
inserting said gene
and said chimeric regulatory region into a plant; and
allowing said
plant to express said gene.
|
Claim 21
A method for inducible expression of a foreign gene in a plant,
comprising:
linking said foreign gene to a regulatory region
comprising a promoter derived from a mannopine synthase gene of
Agrobacterium tumefaciens by deletion to base pair position -138 and an
upstream activating sequence derived from a mannopine synthase gene of
Agrobacterium tumefaciens;
inserting said foreign gene and
said regulatory region in said plant; and
inducing expression of said
foreign gene.
|
Claim 22
A method for inducible expression of a foreign gene in a plant,
comprising:
linking said foreign gene to a regulatory region
comprising
a promoter derived from a mannopine
synthase gene of Agrobacterium tumefaciens,
an upstream
activating sequence derived from a mannopine synthase gene of Agrobacterium
tumefaciens, and
an upstream activating sequence derived from
an octopine synthase gene of Agrobacterium tumefaciens;
inserting said foreign gene and said regulatory region in said plant; and
inducing expression of said foreign gene.
|
|
Claim 24
A method of expressing a gene in a plant, comprising the
steps of:
linking said gene to a chimeric regulatory region
comprising at least three upstream activating sequences derived
from an Agrobacterium tumefaciens octopine synthase gene operably
linked to a promoter derived from an Agrobacterium tumefaciens
mannopine synthase;
inserting said gene and said chimeric regulatory
region into a plant; and
allowing said plant to express said gene.
|
This Canadian patent claims an additional method of expressing a gene in a
plant using a chimeric regulatory region with at least 3 UAS from ocs gene and a
promoter from a mas gene.
|
|
Remarks
|
Related patents were also granted in Russia (RU 2142998 C1), in China (CN
1061376) and in Japan (JP 9505205). An application was filed in Brazil but was
abandoned on March 6, 2001.
|
Note: Patent information was last updated on 22 May 2006. Search terms:
"gene" in abstract and "Purdue" or "Biotechnology research" in applicant. Patent
database: PatentLens in combination with INPADOC.
35S-enhanced mas promoter
Calgene has two granted patents, one in the U.S. and one in Canada on a
promoter region comprising a UAS of the CaMV 35S gene linked to a transcription
initiation region. The patent defines a transcription/translation region of
mas as including the sequences responsible for initiating
transcription and effecting translation of the mas gene derived from
the octopine Ti plasmid of A. tumefaciens. A mas promoter is
included within the transcription/translation region. The region comprises the
TATA box, an upstream activating region (UAS) and a transcription start site. As
described in the patents, the DNA sequences of the region can correspond to:
- from -625 bp upstream the transcriptional start site to +60 bp downstream of
the start site;
- from -300 bp upstream the transcriptional start site to +60 bp downstream of
the start site; or
- from -300 bp to the start site.
The UAS or upstream activating region
(UAR) of the CaMV 35S-enhanced mas promoter
can correspond to a DNA sequence:
- from the -25 position of the CaMV 35S promoter to the -360 position;
- from the -45 position to -90 position; or
- from the -25 position or -45 position to the -168 position.
The mas promoter alone is a weak promoter that is active in all
tissues but which drives gene expression at different levels, being highly
expressed in apical shoots and roots. The promoter of the invention, which
combines the mas promoter with the UAR of the CaMV 35S increases levels
of expression to about 5 to 10-fold higher than the strong double CaMV 35S
promoter, according to the patent.
Some bibliographic data and a summary of the independent claims of the
granted patents are presented in the following table.
35S-enhanced mas promoter - Specific Patent Information
| Patent Number |
Title, Independent Claims and Summary of Claims |
Assignee
|
|
US
5106739
- Earliest priority - 18 April 1989
- Filed - 7 February 1990
- Granted - 21 April 1992
- Expected expiry - 21 April 2009
|
Title - CaMV 35S-enhanced mannopine synthase promoter and
method for using same
Claim 1
A DNA sequence comprising, in the 5' to 3' direction, a
first element linked to a second element, said first element
comprising an upstream activating region of CaMV 35S and said
second element comprising a mannopine synthase transcription
initiation region.
|
Claim 9
A chimeric promoter comprising a CaMV 35S enhanced mannopine
synthase promoter, wherein upon expression of a DNA sequence of interest in a
plant cell under the regulatory control of said promoter, said DNA sequence of
interest is expressible at a level of at least 5-fold higher than expression of
said gene of interest in a plant cell under the regulatory control of a CaMV 35S
enhanced CaMV 35S promoter.
|
Claim 16
A method to increase the expression of an expressible gene of interest under
the regulatory control of a mannopine synthase promoter
comprising the steps of:
providing a CaMV 35S
upstream activating region to the 5' end of a DNA sequence
comprising the mannopine synthase promoter; and
allowing said gene to be expressed.
|
The claims are generally directed to:
- A DNA sequence comprising (in 5' to 3' direction): an UAR of CaMV 35S and a
mas transcription initiation region.
- A chimeric CaMV 35S-enhanced mas promoter regulating the expression of a
gene of interest in a plant. The expression level is at least 5-fold higher than
the expression level achieved under the control of a CaMV 35S-enhanced 35S
promoter.
- A method to increase the expression of a gene of interest by using the
described chimeric promoter.
|
Originally assigned to Calgene
(now owned by
Monsanto)
|
|
CA 1334175
- Earliest priority - 18 April 1989
- Filed - 29 September 1989
- Granted - 31 January 1995
- Expected expiry - 30 January 2012
|
Title - Plant promoter and method for using same
The independent claims are the same as
US
5106739.
|
Note: Patent information was last updated on 28 February 2006. Search terms:
"CaMV" in abstract and "Calgene" in applicant. Patent database: PatentLens in
combination with INPADOC.
Enhancers from the ocs gene
Most of the patents in this group were filed and initially granted to Mycogen
Plant Sciences, Lubrizol Enterprises Inc. or Agrigenetics LP and the
Commonwealth Scientific and Industrial Research Organization (CSIRO) in the
U.S., Europe and Canada. Agrigenetics LP was owned by Mycogen Corp. and Lubrizol
Enterprises Inc. but doing business as Mycogen Plant Sciences. Mycogen Plant
Sciences later changed its name to Mycogen seeds and is now owned by Dow
AgroSciences. For more information about this patent group or licensing, Dow
AgroSciences and CSIRO should be contacted (links are provided in the table on
next page).
Approximate scope of protection
The claims of this group of patents encompasses two enhancer elements. One of
them is a consensus DNA sequence from the non-transcribed region of the promoter
regions of seven opine synthase genes and plant viral
promoters from the cauliflower mosaic virus (CaMV), the figwort mosaic
virus (FMV) and the carnation etched ring virus (CERV). The 20 bp consensus
sequence is: 5'-TGACGTAAGCGATGACGTAA-3'.
The other enhancer element is a 16 bp palindromic sequence isolated from the
5'-untranscribed region of the ocs
gene. The nucleotide sequence is 5'-ACGTAAGCGCTTACGT-3'.
The enhancer elements are linked to any plant-expressible
promoter and any structural gene of interest to form
recombinant DNA molecules. The elements are useful for enhancing the expression
of a nearby plant-expressible gene. The patent describes that "enhancing the
expression" of a gene means that the gene is activated only when the enhancer is
present or gene expression is increased by the enhancer but it does not indicate
how much expression has to be increased for a sequence to be considered as an
enhancer.
The claims of the European and the Canadian patents cover sequences that are
at least 50% homologous to the 16 bp OCS enhancer element. Also, the target
plants for transformation with the recombinant molecules are limited to
monocots. More information of the granted patents are presented
in the table on the following page.
Enhancers from the ocs gene - Specific Patent
Information
| Patent Number |
Title, Independent Claims and Summary of Claims
|
Assignee
|
|
US
5573932
- Earliest priority - 6 February 1987
- Filed - 18 May 1990
- Granted - 12 November 1996
- Expected expiry - 12 November 2013
|
Title - OCS element
Claim 1
A recombinant DNA molecule comprising
a DNA
fragment which fragment is a plant enhancer element capable of being bound by an
OCS transcription factor and displaying the upper band binding pattern
characteristic of the wild-type ocs enhancer in gel retardation assays, said
fragment consisting essentially of a consensus sequence selected from the group
consisting of
5'-T-G-A-C-G-T(C)-A-A-G-C(G)-G(A)-A(C)-T-G(T)-A-C-G-T(C)-A(C)-A(C)-3' and its
reverse sequence, in combination with
(a) a
plant-expressible promoter heterologous to said plant enhancer element wherein
said promoter is placed 3' to said enhancer element, and
(b) a plant-expressible structural gene wherein said gene is placed 3'
to said promoter and under the regulatory control of said enhancer element and
said plant-expressible promoter.
|
Claim 16
A recombinant DNA molecule comprising
a DNA
fragment which fragment is a plant enhancer element capable of being bound by an
OCS transcription factor and displaying the upper band binding pattern
characteristic of the wild-type ocs enhancer in gel retardation assays, wherein
said fragment comprises the sequence 5'-ACGTAAGCGCTTACGT-3' and
its reverse sequence, in combination with
(a) a
plant-expressible promoter heterologous to said plant enhancer element wherein
said promoter is placed 3' to said enhancer element, and
(b) a plant-expressible structural gene wherein said gene is placed 3'
to said promoter and under the regulatory control of said enhancer element and
said plant-expressible promoter.
|
The claims are generally drawn to:
- A recombinant DNA molecule comprising in a 5'-3' direction an OCS enhancer
with an opine synthase enhancer consensus sequence, a plant-expressible promoter
and a plant-expressible structural gene. The structural gene is under the
control of the enhancer and the promoter.
- A recombinant DNA molecule having the same elements as mentioned above, but
the enhancer element has the sequence 5'-ACGTAAGCGCTTACGT-3
|
Mycogen
Plant Sciences, Inc
(now owned by
Dow
AgroSciences)
|
|
US
5710267
- Earliest priority - 6 February 1987
- Filed - 2 June 1995
- Granted - 20 January 1998
- Expected expiry - 20 January 2015
|
Title - OCS element
Claim1
A recombinant DNA molecule comprising
a DNA fragment which fragment is a plant enhancer element capable of
being bound by an OCS transcription factor and displaying the upper band binding
pattern characteristic of the wild-type ocs enhancer in gel retardation assays,
said fragment comprising a consensus sequence selected from the
group consisting of 5'-A-C-G-T-A-A(C)-G-C-G-A(G)-T(A)-G(C)-A-C(T)-G-T(C)-3' and
its reverse sequence, in combination with
(a) a
plant-expressible promoter heterologous to said plant enhancer element wherein
said promoter is placed 3' to said enhancer element, and
(b) a plant-expressible structural gene wherein said gene is placed 3'
to said promoter and under the regulatory control of said enhancer element and
said plant-expressible promoter.
|
Claim 16
A recombinant DNA molecule comprising
a DNA fragment which fragment is a plant enhancer element capable of
being bound by an OCS transcription factor and displaying the upper band binding
pattern characteristic of the wild-type ocs enhancer in gel retardation assays,
wherein said fragment comprises the sequence
5'-ACGTAAGCGCTTACGT-3' and its reverse sequence, in combination with
(a) a plant-expressible promoter heterologous to said plant enhancer element
wherein said promoter is placed 3' to said enhancer element,
and
(b) a plant-expressible structural gene
wherein said gene is placed 3' to said promoter and under the regulatory control
of said enhancer element and said plant-expressible promoter.
|
Claim 18
A recombinant DNA molecule comprising
a DNA fragment which fragment is a plant enhancer element capable of
being bound by an OCS transcription factor and displaying the upper band binding
pattern characteristic of the wild-type ocs enhancer in gel retardation assays,
said fragment comprising at leaset two consensus sequences
selected from the group consisting of
5'-A-C-G-T-A-A(C)-G-C-G-A(G)-T(A)-G(C)-A-C(T)-G-T(C)-3' and its reverse
sequence, in combination with
(a) a
plant-expressible promoter placed 3' to said enhancer element, and
(b) a plant-expressible structural gene wherein said gene
is placed 3' to said promoter and under the regulatory control of said enhancer
element and said plant-expressible promoter.
|
Claim 33
A recombinant DNA molecule comprising
a DNA fragment which fragment is a plant enhancer element capable of
being bound by an OCS transcription factor and displaying the upper band binding
pattern characteristic of the wild-type enhancer in gel retardation assays,
wherein said fragment comprises the sequence 5'-ACGTAAGCGCTACGT-3' and its
reverse sequence, in combination with
(a) a
plant-expressible promoter heterologous to said plant enhancer element wherein
said promoter is placed 3' to said enhancer element, and
(b) a plant-expressible structural gene wherein said gene is placed 3'
to said promoter and under the regulatory control of said enhancer element and
said plant-expressible promoter.
|
This patent is a Continuation of
US
5573932.
The claims are generally directed to three different recombinant DNA
molecules having the same elements as mentioned in
US
5573932. The difference is in the nucleotide sequence of the
enhancer element.
|
Agrigenetics LP (owned by
Mycogen)
&
CSIRO
|
|
US
5837849
- Earliest priority - 6 February 1987
- Filed - 2 June 1995
- Granted - 17 November 1998
- Expected expiry - 17 November 2015
|
Title - OCS element
Claim 1
An isolated DNA fragment which is a plant enhancer capable of activating or
enhancing the transcription level of a plant-expressible gene, said enhancer
element comprising a consensus sequence selected from the group
consisting of
5'-T-G-A-C-G-T(C)-A-A-G-C(G)-G(A)-A(C)-T-G(T)-A-C-G-T(C)-A(C)-A(C)-3' and its
reverse sequence.
|
This patent is a divisional of
US
5573932.
The claim is drawn to an isolated plant enhancer element capable of enhancing
the expression of a gene of interest in a plant having the consensus sequence
5'- TGACGTAAGCGATGACGTAA-3'
|
|
EP
278659 B1
- Earliest priority - 6 February 1987
- Filed - 2 February 1988
- Granted - 24 May 1995
- Expected expiry - 2 February 2008
|
Title - OCS enhancer
Claim 1
A monocotyledonous plant transformed with a recombinant DNA molecule
comprising a plant transcription activating element capable of
activating or enhancing the transcription level of a gene
comprising a sequence having 50% to 100% homology with an
identifying sequence selected from 5'-ACGTAAGCGCTTACGT-3' and its reverse
sequence.
|
Claim 15
A method for enhancing the expression of a plant-expressible gene in plant
tissue comprising the steps of
(a) inserting a
transcription activating element comprising a sequence having
50% to 100% homology to an identifying sequence selected from
5'-ACGTAAGCGCTTACGT-3' and its reverse sequence, in such a way that said
transcription activating element modulates the expression of said gene, and
(b) introducing said recombinant DNA molecule into plant tissue.
|
The claims are directed to:
- A monocot transformed with a recombinant DNA molecule having an enhancer
element with a sequence that is 50 to 100% homologous to the sequence
5'-ACGTAAGCGCTTACGT-3'.
- A method for enhancing the expression of a gene in a monocot by introducing
a recombinant molecule with an enhancer element as described above.
|
Originally assigned to Lubrizol Enterprises, Inc.
and
CSIRO
(Now owned by Agrigenetics
and CSIRO
according to Online Public File Inspection)
|
|
Remarks
|
Patents have lapsed in Austria, Greece, Belgium, France, Gemany, Netherlands,
Spain, Sweden and UK.
Applications were also filed in Japan (JP 63276492 A2) and South Africa (ZA
8800319 A).
|
|
CA 1309365
- Earliest priority - 6 February 1987
- Filed - 5 February 1988
- Granted - 27 October 1992
- Expected expiry - 27 October 2009
|
Title - OCS enhancer
Claim 1
A recombinant DNA molecule comprising a plant transcription activating
element capable of activating or enhancing the transcription level of a gene
comprising a sequence having about 50% to 100% homology with an identifying
sequence selected from 5'-ACGTAAGCGCTTACGT-3' and its reverse sequence.
|
Claim 32
A method for enhancing the expression of a plant-expressible gene in plant
tissue comprising the steps of
(a) inserting a
transcription activating element comprising a sequence having
about 50% to 100% homology to an identifying sequence selected from the group
consisting of 5'-ACGTAAGCGCTTACGT-3' and its reverse sequence into a recombinant
DNA molecule comprising a plant-expressible gene under the control of a promoter
in such a way that said transcription activating element modulates the
expression of said gene, and
(b) introducing said
recombinant DNA molecule into plant tissue.
|
The claims are the same as the claims of the European patent
EP
278659 B1 except that transformation of a plant by the recombinant
molecule is not limited to monocots.
|
Lubrizol Enterprises, Inc. (It is most likely part of
Mycogen Seeds
now)
and
CSIRO
|
Note: Patent information was last updated on 22 May 2006. Search terms: "ocs
element" in abstract. Patent database: PatentLens in combination with INPADOC.
Plant Ubiquitin promoter (Ubi)
Scientific aspects
Ubiquitin is a protein found in eukaryotic cells and its sequence is highly
conserved among organisms as diverse as humans and the fruit fly. The protein is
implicated in processes such as protein turnover, chromatin structure, cell
cycle control, DNA repair, and response to heat shock and other stresses.
In 1992, Christensen et al. identified two out of the 8 to 10 loci
encoding ubiquitin in maize. Both characterized genes, Ubi-1 and
Ubi-2, contain an open reading frame of 1599 bp arranged as seven
tandem, head-to-tail repeats of 228 bp.
The regulatory region which controls the expression of the Ubi-1
gene of maize extends from -899 bp 5' of the transcription start site (+1) to
about 1093 bp 3' of the transcription start site.
This sequence of approximately 2 kb comprises:
- a TATA box sequence located at -30,
- two overlapping sequences that are similar to the consensus heat shock
element found in heat-inducible genes, located at the -214 and -204 position
from the transcription start site,
- an 83 bp untranslated exon sequence 3' of and adjacent to the transcription
start site and
- an intron of around 1 kb, which extends from 84 to 1093 position.
The heat shock elements of the regulatory region enhance the expression of
the ubiquitin protein in response to temperature stress.
IP issues
Mycogen Plant Science (now owned by Dow Agro Sciences) and
Monsanto PLC are entities that have patents
and patent applications on the plant ubiquitin regulatory system. The
Mycogen patents evaluated in this report were granted in:
- the United States (4 patents)
- Europe (1 patent), and
- Canada (1 patent)
Monsanto has granted patents in the United States and
Australia. Related patent applications were also filed in Europe and Canada.
Prodigene owns a United States patent and also filed a
patent application in Australia.
How is promoter defined?
The description of a eukaryotic promoter in both Mycogen
patents and Monsanto patent applications includes functional
and structural components. That is, a promoter is defined as the region upstream
of a gene containing the binding site for RNA polymerase II that initiates
transcription of the DNA. It contains a TATA box, a CAAT box or an AGGA box, and
the CAP site. The inventors also acknowledge that there are ancillary regulatory
sequences or regions that are part of a regulatory system in general. Those
regions include enhancers and upstream activating sequences.
One of the regulatory elements set out by the inventors of the
Mycogen patents is the heat shock element,
which transiently enhances the level of downstream gene expression in response
to sudden temperature elevation.
Thus, the inventions described in the patents do not refer to a "simple"
ubiquitin promoter, but to a ubiquitin regulatory system that
includes the promoter as described above and the additional elements
that participate in the modulating gene expression.
In the Monsanto patents, the ubiquitin regulatory
system is modified by excising the
heat shock elements. While In the Prodigene
patent, the ubiquitin regulatory system is modified by making
the two overlapping heat shock elements adjacent.
Patents owned by Mycogen Plant Science (probably currently
controlled by Dow AgroSciences)
|
Patent number
|
Title, Summary of Claims and Independent
Claims
|
Assignee
|
|
US
5510474
- Earliest priority - 17 May 1988
- Filed - 25 August 1994
- Granted - 23 April 1996
- Expected expiry - 23 April 2013
|
Title - Plant ubiquitin promoter system
|
Claim 1
An isolated DNA fragment, useful in effecting
expression in both monocots and dicots of coding sequences placed 3' to said
fragment, wherein said DNA is approximately 2 kb in length, and said DNA
fragment further comprises, in the following order beginning
with the 5' most element and proceeding toward the 3' terminus of said DNA
fragment:
(a) two heat shock elements, which overlap;
(b) a promoter
comprising a transcription start site;
(c) an intron
of about 1 kb in length; and
(d) a translation start site; wherein
said DNA fragment comprising said elements (a)-(d) regulates
gene expression in both dicots and monocots, and wherein said DNA fragment
comprises the nucleotide sequence shown from position -899 to
1092 of the maize ubiquitin sequence listed in
FIG.
2.
|
Independent claim 1 is drawn to:
- An isolated DNA fragment from -899 to +1092 of the 5' flanking region of a
maize ubiquitin sequence comprising from 5' to 3': two overlapping heat shock
elements, a promoter with a transcription start site, an intron of about 1 kb,
and a translation start site.
- The DNA fragment has to be capable of regulating gene expression (of any
gene) in both dicots and monocots; and
- must include a portion of the sequence shown in Figure 2 of the patent
(i.e., the sequence from position -899 to 1092).
|
Mycogen
Plant Sciences
|
|
US
5614399
- Earliest priority - 17 May 1988
- Filed - 5 June 1995
- Granted - 25 March 1997
- Expected expiry -25 March 2014
|
Title - Plant ubiquitin promoter system
|
Claim 1
A method for selective heat shock induced enhancement of the constitutive
expression of a structural gene in a plant cell comprising the
steps of:
(a) transforming said plant cell with a DNA construct
comprising an approximately 2 kb plant ubiquitin regulatory
region operably joined to a DNA sequence of interest, wherein said plant
ubiquitin regulatory region is from a plant ubiquitin gene and
comprises at least one heat shock element, a promoter, a
transcription start site, and an intron; and
(b) selectively applying
stress conditions of high temperature to said transformed plant cell thereby
inducing enhancement in expression of said DNA sequence of interest.
|
This patent is a divisional of
US
5510474. The claims are drawn to:
A method for rendering a constitutively expressed structural gene of a plant
cell heat shock inducible by transforming the plant cell with a DNA construct
which comprises a plant ubiquitin regulatory region and selectively applying
high temperature to the transformed cell.
The plant ubiquitin regulatory region comprises:
- at least one heat shock element
- a promoter
- a transcription start site
- and an intron
The plant can be any type but the gene must be a structural gene (defined in
the patent as one which encodes a protein, polypeptide or portion thereof) and
so the claim does not cover constructs for expressing siRNAs.
The term "high temperature" is not defined but the patent describes that heat
shock can occur at 40° C.
The patent discloses a single type of ubiquitin regulatory system - the
sequence from a single maize ubiquitin gene - though the claims encompass
ubiquitin regulatory systems from any kind of plant.
|
|
US
6020190
- Earliest priority - 17 May 1988
- Filed - 18 November 1996
- Granted - 1 February 2000
- Expected expiry - 17 May 2008
|
Title - Plant ubiquitin promoter system
|
Claim 1
A DNA construct comprising:
(a) a DNA sequence no larger than 2 kb, said DNA sequence
comprising a plant ubiquitin regulatory system, wherein said
regulatory system contains a heat shock element and an intron, said intron being
located at 3' to said heat shock element, and
(b) a plant-expressible
structural gene wherein said structural gene is placed under the regulatory
control of said plant ubiquitin regulatory system.
|
This patent is a divisional of
US
5614399. The claims generally relate to:
- A DNA construct having a plant ubiquitin regulatory sequence no larger than
2 kb that contains an intron located 3' of a heat shock element, and a
plant-expressible structural gene under the control of the ubiquitin regulatory
sequence.
Although the claim recites that the construct must contain a plant
expressible gene this does not necessarily mean that the gene is from a plant.
However, the claim does require that the gene is a structural gene (e.g.,
must encode a protein, polypeptide or portion thereof).
The patent describes that placing a structural gene under the regulatory
control of a promoter or a regulatory element "means positioning the structural
gene such that the expression of the gene is controlled by these sequences. In
general, promoters are found positioned 5' (upstream) to the genes that they
control."
|
|
US
6054574
- Earliest priority - 17 May 1988
- Filed - 9 June 1998
- Granted - 25 April 2000
- Expected expiry - 17 May 2008
|
Title - Plant ubiquitin promoter system
|
Claim 1
A DNA fragment approximately 2 kb in length, said DNA fragment
comprising a plant ubiquitin regulatory system, wherein said
regulatory system contains a promoter comprising
a
transcription start site,
one or more heat shock elements positioned
5' to said transcription start site, and
an intron positioned 3' to
said transcription start site,
wherein said regulatory system is capable
of regulating constitutive and inducible gene expression in both dicots and
monocots such that the level of said constitutive gene expression in monocots is
about one-third that obtained in said inducible gene expression in monocots.
|
|
Claim 9
A recombinant DNA construct comprising:
a. A DNA fragment approximately 2 kb in length, said DNA fragment
comprising a plant ubiquitin regulatory system, wherein said
plant ubiquitin regulatory system contains a promoter
comprising
a transcription start site,
one more heat shock elements positioned 5' to said transcription start
site,
a translational start site, and
an intron
positioned 3' to said transcription start site and 5' to said translational
start site, wherein said plant ubiquitin regulatory system is capable of
regulating constitutive and inducible gene expression in both dicots and
monocots such that said constitutive gene expression in monocots is at a level
about one-third that obtained in said inducible gene expression in monocots, and
b. a plant-expressible heterologous structural gene positioned 3' to said
plant ubiquitin regulatory system and a polyadenylation signal positioned 3' to
said structural gene, wherein said heterologous gene is placed under the
regulatory control of said plant ubiquitin regulatory system.
|
|
Claim 18
A DNA fragment, useful in effecting expression in both monocots and dicots of
coding sequences placed 3' to said fragment, wherein said DNA is isolated or
incorporated into a larger piece of DNA but in a position other than in the 5'
sequence of a plant ubiquitin gene, is approximately 2 kb in length, and said
DNA fragment further comprises, in the following order
beginning with the 5' most element and proceeding toward the 3' terminus of said
DNA fragment:
(a) one or more heat shock elements, which elements may or may not be
overlapping;
(b) a promoter comprising a transcription start site; and
(c) an intron of about 1 kb in length; and wherein said DNA fragment
comprising said elements (a)-(c) is capable of regulating gene expression in
both dicots and monocots.
|
This patent is a divisional of
US
6020190. The claims generally relate to:
- A DNA fragment of approximately 2 kb comprising a plant ubiquitin regulatory
system comprising: one or more heat shock elements 5' to a promoter with a
transcription start site and an intron 3' to the transcription start site. The
system can be used to regulate constitutive and inducible expression for both
dicots and monocots; however, constitutive expression must be 1/3 the level of
heat-shock inducible expression in monocots.
- A recombinant DNA construct having a plant ubiquitin regulatory system as
described and a plant expressible heterologous structural gene under control of
the regulatory region which also includes a polyadenylation sequence 3' of the
structural gene.
- A DNA fragment of approximately 2kb as described above which can be part of
a larger sequence so long as it is not 5' of a plant ubiquitin gene. The level
of heat-shock inducibility in monocots is not an element of this claim.
A particular level of heat shock induction in dicots is not an element of any
of the claims.
The ubiquitin regulatory system can be from any plant although the patent
does not describe any sequences other than those from maize.
|
|
CA 1339684
- Earliest priority - 17 May 1988
- Filed - 16 May 1989
- Granted - 24 February 1998
- Expected expiry - 24 February 2015
|
Title - Plant ubiquitin promoter system
|
Claim 1
A DNA fragment approximately 2kb in length, said DNA fragment
comprising a plant ubiquitin regulatory system, wherein said
regulatory system contains:
a) overlapping heat shock elements
and
b) an intron, and
wherein said regulatory system is capable of
regulating constitutive and inducible gene expression in both dicots and
monocots.
|
|
Claim 11
A recombinant DNA construct
comprising:
a) a DNA fragment approximately 2kb in
length, said DNA fragment comprising a plant ubiquitin regulatory system,
wherein said regulatory system contains:
1. overlapping heat shock
elements and
2. an intron; and
wherein said regulatory system
is capable of regulating constitutive and inducible gene expression in both
dicots and monocots, and
b) a plant-expressible heterologous structural gene wherein said
heterologous structural gene is placed under the regulatory control of said
plant ubiquitin regulatory system.
|
|
Claim 23
A method for the constitutive expression of a structural gene and the
selected stress-induced enhancement in expression of said structural gene in a
plant cell comprising the steps of:
a) transforming
said plant cell with a DNA construct comprising an
approximately 2kb plant ubiquitin regulatory system, wherein is found a heat
shock element and an intron, and
b) a plant-expressible structural gene
that is under the regulatory control of said plant regulatory system, and
c) selectively applying stress conditions of high temperature to said
transformed plant cell thereby inducing enhancement in expression of said
structural gene.
|
The claims are generally drawn to:
- plant ubiquitin regulatory system of around 2 kb capable of constitutive
and inducible regulation of gene expression in both monocots and dicots. The
system contains an overlapping heat shock elements and intron.
- A recombinant DNA construct having a plant ubiquitin regulatory system as
described and a plant expressible structural gene under control of the
regulatory region.
- A method for both constitutive and stress-inducible enhanced expression of
a structural gene in a plant cell. The transformed cell with the plant ubiquitin
system is subjected to high temperature to induce enhancement in gene
expression.
The construct used in method claim 23 does not require overlapping heat shock
elements but encompasses constructs with a single heat shock element.
The claims don't require a particular level of induction after heat shock.
As with the above patents, the ubiquitin regulatory system could be from any
plant so long as that system comprises a heat shock element.
|
|
EP
342926 B1
- Earliest priority - 17 May 1988
- Filed - 16 May 1989
- Granted - 28 September 1994
- Expected expiry - 16 May 2009
|
Title - Plant ubiquitin promoter system
|
Claim 1
A DNA sequence no larger than 2 kb, said DNA
sequence comprising a plant ubiquitin regulatory system,
wherein said regulatory system contains:
a) a heat shock element
and
b) an intron.
|
|
Claim 10
A DNA construct comprising:
a) a DNA sequence no larger than 2 kb, said DNA sequence comprising a
plant ubiquitin regulatory system, wherein said regulatory system contains:
1. a heat shock element, and
2. an intron; and
b) a plant-expressible structural gene wherein said structural gene is placed
under the regulatory control of said plant ubiquitin regulatory system.
|
|
Claim 12
A method for the constitutive expression of a
structural gene and the selected stress-induced enhancement in expression of
said structural gene in a plant cell comprising the steps of:
a) transforming said plant cell with a DNA construct
comprising:
i. a plant ubiquitin regulatory
system, wherein is found a heat shock element and an intron, and
ii. a plant-expressible structural gene that is under the regulatory control of
said plant ubiquitin regulatory system, and
b) selectively applying stress conditions to said transformed plant cell
thereby inducing enhancement in expression of said structural gene.
|
|
|
Remarks
|
The European patent converted to and in force in Belgium (BE), France (FR),
Germany (DE), Great Britain (GB), Italy (IT), Spain (ES) and the Netherlands
(NL). A patent (JP 3509855) was also granted in Japan.
|
Note: Patent information was last updated on 23 May 2006. Search terms:
"ubiquitin" in abstract and "Mycogen" in applicant. Patent database: PatentLens
in combination with INPADOC.
Patents and applications relating to the ubiquitin regulatory region owned
by Monsanto
|
Patent/application number
|
Title, Independent Claims and
Summary of Claims
|
Assignee
|
|
US
6878818
- Earliest priority - 9 September 1999
- Filed - 12 June 2000
- Granted - 12 April 2005
- Expected expiry - 9 September 2019
|
Title - Modified ubiquitin regulatory system
|
Claim 1
An isolated DNA sequence comprising a ubiquitin regulatory
system lacking heat shock elements wherein the ubiquitin regulatory system
comprises the nucleotide sequence according to
SEQ.ID.NO.
8.
|
The claims are to:
- DNA sequences which comprise a specific nucleotide sequence
(SEQ.ID.NO.
8), a 2033 bp sequence which includes the promoter, the 83 bp untranslated
first exon and the intron of the maize Ubi-1 gene.
- Dependent claims are drawn to host cells, transgenic plants (of any kind),
and seeds comprising the sequences, as well as constructs comprising the
sequence operably linked to a plant-expressible structural coding sequence and
vectors comprising the constructs.
Claim 1 requires that the DNA sequence has at least the 2033 bp sequence but
allows for the inclusion of additional sequences - such as the remainder of the
coding region of the Ubi-1 gene or heterologous sequences not usually
found in association with the Ubi-1 promoter and intron. However, the
heat shock elements cannot be included.
Because the claims do require
SEQ.ID.NO.
8 as an element of the sequence, a regulatory system which included portions
of the promoter and intron sequence required for expression, but which deleted
other parts of the sequence would not literally be covered by these claims.
|
Monsanto
UK Ltd
|
|
AU
769567 B2
- Earliest priority - 9 September 1999
- Filed - 7 September 2000
- Granted - 29 January 2004
- Expected expiry - 7 September 2020
|
Title - Modified ubiquitin regulatory system
|
Claim 1
A DNA sequence comprising a
ubiquitin regulatory system lacking heat shock elements wherein the ubiquitin
regulatory system comprises the nucleotide sequence according
to
SEQ.ID.NO.
8.
|
Claim 1 of the Australian patent is identical to that of the US patent.
|
|
EP 1210446 A1
- Earliest priority - 9 September 1999
- Filed - 7 September 2000
- Status - Pending
- Expected expiry - N/A
|
Title - Modified ubiquitin regulatory system
|
Claim 1
A DNA sequence comprising an ubiquitin regulatory system
lacking heatshock elements.
|
|
Claim 2
A DNA sequence comprising an ubiquitin regulatory system
lacking heatshock elements.
|
These independent claims are broader than those in the US patent since the
exact base composition of the ubiquitin regulatory system is not an element of
the claims and the system can be from any organism.
The application discloses that a ubiquitin regulatory system which lacks heat
shock elements includes a promoter, a transcription start site, an intron and
optionally a translation start site. Only the regulatory sequences of one maize
ubiquitine gene is disclosed.
|
|
CA 2384517
- Earliest priority - 9 September 1999
- Filed - 7 September 2000
- Status- Pending
- Expected expiry - N/A
|
Title - Modified ubiquitin regulatory system
|
Claim 1
A DNA sequence comprising a
ubiquitin regulatory system lacking heat shock elements wherein the ubiquitin
regulatory system comprises the nucleotide sequence according
to
SEQ.ID.NO.
8.
|
The claims of this Canadian application are exactly the same as
AU
769567 B2.
|
Note: Patent information was last updated on 23 May 2006. Search terms:
"ubiquitin" in abstract and "Monsanto" in applicant. Patent database: PatentLens
and esp@cenet in combination with INPADOC.
Patent owned by Prodigene
|
Patent/application number
|
Title, Independent Claims and
Summary of Claims
|
Assignee
|
|
US
6977325
- Earliest priority - 9 Jun 2000
- Filed -28 Feb 2002
- Granted- 20 Dec 2005
- Expected expiry - 16 Jun 2020 (adjusted)
|
Title - Plant promoter sequences and methods of
use for same
Claim 1
An engineered ubiquitin promoter sequence capable of directing expression of
a nucleotide sequence in a plant cell, said engineered ubiquitin promoter
sequence comprising: a heat shock region, wherein said heat shock region has the
sequence as set forth in
SEQ
ID NO: 4.
|
Claim 2
A method for causing expression of a heterologous structural gene or open
reading frame in a plant cell, said method comprising: introducing to a plant
cell an expression construct comprising an engineered ubiquitin promoter
sequence operably linked to said heterologous structural gene or open reading
frame, wherein said engineered ubiquitin promoter sequence comprises a heat
shock region, wherein said heat shock region has the sequence as set forth in
SEQ
ID NO: 4.
|
|
Prodigene Inc.
|
|
Remarks
|
The patent application filed in Australia (AU 200175433)
lapsed. The related PCT application is
WO
2001/094394.
|
|
Note: Patent information was last updated on 30 October 2006. Search terms:
"ubiquitin" in abstract and "Prodigene" in applicant. Patent database:
PatentLens and in combination with INPADOC.
Ubiquitin promoters from rice and sugarcane
Apart from patents on the ubiquitin regulatory system from maize, there are
also patents specifically claiming ubiquitin promoters from rice and sugarcane.
The Board of Supervisors of Louisiana State University and Agricultural and
Mechanical College holds a United States patent
(US
6528701) on rice ubiquitin promoters. This patent claims a rice
ubiquitin promoter and a method for using it in controlling gene expression in
plants. It shows in the description that the rice ubiquitin promoter is stronger
than the maize ubiquitin promoter in driving gene expression in rice.
United States of America as represented by the Secretory of Agriculture &
University of Hawaii has three United States patents
(US
6706948,
US
6686513 and
US
6638766) on sugarcane ubiquitin promoters (UBI4 and UBI9).
rice ubiquitin promoters
|
Title, Independent Claims and
Summary of Claims
|
Assignee
|
|
US
6528701
- Earliest priority - 29 Feb 2000
- Filed - 29 Feb 2000
- Granted - 04 Mar 2003
- Expected expiry - 29 Feb 2020
|
Title - Rice ubiquitin-derived promoters
Claim 1
An isolated nucleotide sequence comprising a rice ubiquitin promoter capable
of controlling constitutive expression of a nucleic acid encoding a polypeptide,
wherein said nucleotide sequence comprises at least a portion of
SEQ
ID NO:3 which is upstream of position 2785 and wherein said portion retains
promoter activity.
|
The claims of this patent are generally drawn to:
- An isolated nucleotide sequence comprising a rice ubiquitin promoter
(upstream of position 2785 of SEQ ID NO:3) , and
- A method of expressing a heterologous nucleic acid sequence encoding a
polypeptide in a plant or in its progeny under the control of the rice ubiquitin
promoter (claim 6).
|
Board of Supervisors of Louisiana State University and
Agricultural and Mechanical College
|
Note: Patent information was last updated on 31 October 2006. Search terms:
"Ubiquitin promoter" in abstract. Patent database: PatentLens in combination
with INPADOC.
Patents and applications on sugarcane ubiquitine promoters
|
Patent/application number
|
Title, Independent Claims and
Summary of Claims
|
Assignee
|
|
US
6706948
- Earliest priority - 19 Mar 1998
- Filed - 17 Mar 1999
- Granted - 16 Mar 2004
- Expected expiry - 17 Mar 2019
|
Title - Sugarcane UBI9 gene promoter and methods of use
thereof
Claim 1
A substantially purified nucleic acid sequence comprising a nucleotide
sequence selected from the group consisting of
SEQ
ID NO:7, the complement of
SEQ
ID NO:7,
SEQ
ID NO:10, and the complement of
SEQ
ID NO:10.
|
Claim 4
A substantially purified nucleic acid sequence comprising the HindIII/XbaI
fragment isolated from plasmid pubi9-GUS contained in Escherichia coli cells
deposited as NRRLB-30116.
|
Claim 5
A transgenic plant cell comprising a nucleic acid sequence comprising a
double-stranded nucleotide sequence listed as
SEQ
ID NO:10, wherein said nucleotide sequence is operably linked to a nucleic
acid sequence of interest.
|
Claim 6
A method for expressing a nucleic acid sequence of interest in a plant cell,
comprising:
a) providing:
i) a plant cell;
ii) a nucleic acid sequence of interest; and
iii) a nucleotide sequence selected from the group consisting of
SEQ
ID NO:10, and the complement of
SEQ
ID NO:10;
b) operably linking said nucleic acid sequence of interest to said
nucleotide sequence to produce a transgene; and
c) introducing said transgene into said plant cell to produce a
transgenic plant cell under conditions such that said nucleic acid sequence of
interest is expressed in said transgenic plant cell.
|
Claim 7
A method for expressing a nucleic acid sequence of interest in a plant cell,
comprising:
a) providing:
i) a plant cell;
ii) a nucleic acid sequence of interest; and
iii) a promoter comprising
SEQ
ID NO:10;
b) operably linking said nucleic acid sequence of interest to said
promoter to produce a transgene;
c) introducing said transgene into said plant cell to produce a
transgenic plant cell under conditions such that said nucleic acid sequence of
interest is expressed in said transgenic plant cell, and
d) identifying said transgenic plant cell.
|
The claims of this patent are directed to:
|
United States of America as represented by the
Secretory of Agriculture & University of Hawaii
|
|
US
6686513
- Earliest priority - 19 Mar 1998
- Filed - 20 Oct 2000
- Granted - 03 Feb 2004
- Expected expiry - 17 Mar 2019
|
Title - Sugarcane ubi9 gene promoter sequence and methods of
use thereof
Claim 1
A transgenic plant cell comprising a promoter comprising
SEQ
ID NO:10, wherein said promoter is operably linked to a nucleic acid
sequence of interest, and said plant cell is selected from sugar cane, tobacco,
sorghum, pineapple, rice, maize, tomato, soybean, banana, and garlic.
|
Claim 6
A method for expressing a nucleic acid sequence of interest in a plant cell
selected from sugar cane, tobacco, sorghum, pineapple, rice, maize, tomato,
soybean, banana, and garlic, comprising:
a) providing:
i) said plant cell;
ii) a nucleic acid sequence of interest; and
iii) a promoter comprising
SEQ
ID NO: 10;
b) operably linking said nucleic acid sequence of interest to said
promoter to produce a transgene; and
c) introducing said transgene into said plant cell to produce a
transgenic plant cell under conditions such that said nucleic acid sequence of
interest is expressed in said transgenic plant cell.
|
This patent is a Continuation in part of
US
6706948.
|
|
US
6638766
- Earliest priority - 19 Mar 1998
- Filed - 24 May 2001
- Granted - 28 Oct 2003
- Expected expiry - 4 Jun 2019 (adjusted)
|
Title - Promoter of the sugarcane UBI4 gene
Claim 1
A substantially purified nucleic acid sequence comprising the nucleotide
sequence selected from the group consisting of SEQ ID NO:7 and the complement
thereof.
|
Claim 4
A transgenic plant cell comprising a nucleic acid sequence comprising the
double-stranded nucleotide sequence listed as
SEQ
ID NO:7, wherein said nucleotide sequence is operably linked to a nucleic
acid sequence of interest.
|
This patent is a Continuation of
US
6706948.
|
|
Remarks
|
Related patent applications were also filed in Australia (AU 3192799
A1), Europe (EP 1063880 A1), Canada
(CA 2324520) and Israel (IL 138580). However,
the applications in Australia, Europe and Canada all lapsed.
|
Note: Patent information was last updated on 31 October 2006. Search terms:
"polyubiquitin" in abstract. Patent database: PatentLens in combination with
INPADOC.
Plant Actin promoters
Scientific aspects
In the quest to improve the expression of foreign genes in transformed
cereals, several promoter regions originating from monocot plants have been
tested. In 1990, McElroy and colleagues from Cornell University reported that
the 5' region of the rice actin 1 (Act-1) gene successfully directed
the expression of a reporter gene in transformed rice protoplasts. Since then,
the promoter of the rice Act-1 gene has been used as a strong
constitutive promoter driving the expression of genes of interest in monocots.
Actin is a fundamental component of the cell cytoskeleton present in all
tissues. Cell shape determination, cell division, organelle movement and
extension growth are all believed to involve actin proteins.
The Act-1 gene from rice has a short 5' non-coding exon, separated
by a 447 bp intron (intron 1) from the first coding exon. The presence of the
first intron of the gene has proved to be fundamental for the efficient gene
expression from the Act-1 promoter. The region is active in almost all
plant tissues except sporophytic and gametophytic pollen tissues.
All the necessary cis-acting regulatory elements for the activity of
the Act-1 promoter are in the region 834 bp upstream of the
transcription start site. Apart from the TATA box, there are two regions that
play a role in the regulation of the constitutive expression of the promoter. A
poly (dA-dT) element of 38 bp between -245 and -152 position is the binding
sequence for a trans-acting protein that works as a positive regulator
of Act-1 promoter activity. A region between -300 and -260 positions
contains CCCAA pentamer repeats that appear to be involved in negative
regulation of promoter activity. Its action seems to be tissue-specific,
particularly in roots.
The portion of the rice Act-1 gene used in vectors for monocot
transformation normally contains:
- approximately 1 kb of regulatory sequences located 5' of the
transcribedregion,
- the 5' non-coding exon 1,
- the intron 1, and
- the coding exon 2 of the Act-1 gene.
The regulatory region of rice Act-1 gene has been successfully used for
expressing diverse genes of interest after transformation of cereals, i.e.
maize, rice, barley, wheat and rice.
IP issues
1. Promoter region derived from the rice Act-1 gene
The rice Act-1 promoter region is protected under a
U.S. patent granted to the Cornell
Research Foundation, Inc. In the description of the invention, the
definition of a promoter regulatory region includes more sequence than the
minimal elements of a promoter region: TATA box, AGGA box and transcription
start site. The claimed promoter region encompasses sequences located 5' to the
translational start site that are essential for conferring a high level of
expression of the gene under its control.
Approximate scope of protection
The claimed isolated nucleotide sequence of the Act-1 promoter construct
extends 2.1 kb in the 5' direction from the translation initiation
codon. The construct contains:
- 1.3 kb of 5' untranscribed sequence
- the 5' transcribed but untranslated exon
- the 5' intron, and
- a part of the first coding exon of the gene.
From the description of the invention it can be concluded that the presence
of the 5' intron is vital for the efficient performance of the
promoter construct.
Specific patent information
| Patent number |
Title, Independent Claims and Summary of
Claims |
Assignee
|
|
US
5641876
- Earliest priority - 5 January 1990
- Filed - 27 October 1993
- Granted - 24 June 1997
- Expected expiry - 24 June 2014
|
Title - Rice actin gene and promoter
Claim 1
An isolated nucleic acid molecule encoding a promoter region from rice actin
1 gene.
|
Claim 2
An isolated nucleic acid molecule encoding a promoter region from rice actin
1 gene wherein said nucleic acid molecule has a nucleotide sequence as shown in
nucleotides 1-2180 of
SEQ
ID NO:5.
|
The claims are directed to:
- An isolated DNA sequence encoding a promoter from rice Act-1 gene.
The nucleotide sequence of 2.18 kb is provided.
|
Cornell Research
Foundation, Inc.
|
|
Remarks
|
The PCT application
WO
91/09948 was not converted into national applications.
The patent application in Australia ( AU 71827/91) lapsed.
|
Note: Patent information was last updated on 22 May 2006. Search terms:
"actin" in abstract and "cornell" in applicant. Patent database: PatentLens in
combination with INPADOC.
2. Elements of the 5' region of the rice Act-1 gene.
As mentioned before, apart from the promoter region itself (5' upstream of
the transcription start site) of the rice Act-1 gene, there are
elements of the 5' region of the gene that play an important regulatory role
in driving expression of a downstream linked gene.
The regulatory elements are used in a modular way, in conjunction with other
promoters or regulatory transcriptional regions. Patents and patent applications
directed to the use of rice Act-1 derived elements have claims that
include:
- the intron 1 of the rice Act-1 gene, and
- the exon 1 of the same gene.
A. Intron 1
- as part of a wound-inducible promoter
construct for monocots. A
U.S. patent granted to Cornell
Research Foundation protects:
- the use of a 5' intron sequence of the rice Act-1 gene linked to a
wound-inducible potato proteinase inhibitor II gene promoter to form a promoter
region and
- a construct, a monocot plant and a rice plant containing a foreign gene of
interest under the control of the mentioned promoter region.
- as part of a chimeric promoter for driving the expression
of a gene of interest in transformed monocots. There is a
European patent application and an
Australian patent application filed by
Rhone Poulenc disclosing a chimeric regulatory region
comprising the first intron of the rice Act-1 gene and a promoter derived from
the maize histone H3C4 gene. In some of the filed independent claims, the gene
of interest encodes an optimized transit peptide-calcium-dependent protein
kinase (OTP/CP4).
B. Exon 1
Granted Australian, European and
U.S. patents assigned to the
Max-Planck Institute (Gottingen, DE) are directed to a modular
promoter construct for plant transformation having a DNA sequence from
exon 1 of the rice Act-1 gene regulatory region and a
promoter. The promoter of the modular construct is
any but the rice Act-1 gene promoter. Derivatives of
the modular promoter construct are also covered.
The U.S. patent also claims a vector for
plant cell transformation containing the modular promoter construct, a
transformed plant and its descendants, and a method for preparing plants with an
elevated gene expression level by transforming the plant with a vector
comprising the claimed modular promoter construct.
Specific patent information
|
Patent number
|
Title, Independent Claims and Summary
of Claims
|
Assignee
|
|
US
5684239
- Earliest priority - 5 January 1990
- Filed - 9 September 1994
- Granted - 4 November 1997
- Expected expiry - 4 November 2014
|
Title - Monocot having dicot wound-inducible promoter
Claim 1
A wound inducible promoter construct for use in monocotyledonous plants
consisting essentially of, in 5' to 3' order, a potato
proteinase inhibitor II gene promoter and a 5' intron of rice actin 1 gene
promoter.
|
Claim 2
A nucleic acid construct comprising: a wound inducible promoter construct
consisting essentially of, in 5' to 3' order, a potato
proteinase inhibitor II gene promoter and a 5' intron of rice actin 1 gene
promoter; and a foreign gene of interest under regulatory control of said wound
inducible promoter construct.
|
Claim 3
A monocotyledonous plant comprising: a wound inducible
promoter construct consisting essentially of, in 5' to 3' order, a potato
proteinase inhibitor II gene promoter and a 5' intron of rice actin 1 gene
promoter; and a foreign gene of interest under regulatory control of said wound
inducible promoter construct.
|
Claim 4
A rice plant comprising: a wound inducible promoter
construct consisting essentially of, in 5' to 3' order, a potato proteinase
inhibitor II gene promoter and a 5' intron of rice actin 1 gene promoter; and a
foreign gene of interest under regulatory control of said wound inducible
promoter construct.
|
|
Cornell Research
Foundation, Inc.
|
|
Remarks
|
The related European application EP 666921 was withdrawn.
Application also filed in Japan (JP 7503126 T2).
|
|
US
6750378
- Earliest priority - 24 December 1997
- Filed - 10 March 1998
- Granted - 15 June 2004
- Expected expiry - 24 December 2014
|
Title - Maize H3C4 promoter combined with the first intron
of rice actin, chimeric gene comprising it and transformed plant
Claim 1
An isolated DNA sequence comprising, in the direction of
transcription, a fragment of the sequence of the maize H3C4 promoter wherein
said fragment of the sequence of the maize H3C4 promoter has the sequence shown
in
SEQ
ID NO: 1 and a fragment of the sequence of the first intron of rice actin
wherein said fragment of the sequence of the first intron of rice actin has the
sequence shown in
SEQ
ID NO: 2.
|
Claim 2
An isolated DNA sequence comprising the DNA sequence shown in
SEQ
ID NO: 3.
|
|
Rhone-Poulenc Agrochimie
(then named Aventis CropScience SA
and now
Bayer
cropScience )
|
|
US
2002/104117 A1
- Earliest priority - 24 December 1997
- Filed - 10 March 1998
- Granted - Pending
- Expected expiry - N/A
|
Title - Maize H3C4 promoter combined with the first intron
of rice actin, chimeric gene comprising it and transformed plant
This patent application is a Continuation of
US
6750378.
Claim 1
DNA sequence, a 5' regulatory element allowing the expression of a
heterologous gene in a plant cell from a monocotyledonous plant, characterized
in that it comprises, in the direction of transcription, a first DNA sequence,
which is a functional fragment of the sequence of the maize H3C4 promoter, and a
second DNA sequence, which is a functional fragment of the sequence of the first
intron of rice actin.
|
Claim 8
DNA sequence, a 5' regulatory element allowing the expression of a
heterologous gene in a plant cell from a monocotyledonous plant, characterized
in that it comprises the DNA sequence represented by the
sequence identifier No. 3
(SEQ
ID NO: 3) or a sequence homologous to the said sequence.
|
Claim 21
DNA sequence encoding a fusion protein OTP/CP4.
|
Claim 22
Fusion protein OPT/CP4.
|
Claim 23
Chimeric gene characterized in that it comprises, in the
direction of transcription, an appropriate 5' regulatory sequence for ensuring
the expression of a coding sequence in plants, functionally linked to a sequence
encoding a fusion protein OTP/CP4, optionally linked to a 3' regulatory
sequence.
|
|
|
US
2004/199944 A1
- Earliest priority - 24 December 1997
- Filed - 16 January 2004
- Granted - Pending
- Expected expiry - N/A
|
Title - Maize H3C4 promoter combined with the first intron
of rice actin, chimeric gene comprising it and transformed plant
This patent application is a Continuation of
US
6750378.
Claim 2
Fusion protein OTP/CP4.
|
Claim 4
An isolated DNA sequence comprising, in the direction of
transcription, a functional fragment of the sequence of the maize H3C4 promoter
and a functional fragment of the first intron of rice actin.
|
| Claim 9
An isolated DNA sequence comprising a sequence homologous to
SEQ
ID NO: 3.
|
Claim 22
An isolated DNA sequence encoding a fusion protein OTP/CP4.
|
Claim 23
An expression cassette comprising in the direction of
transcription, a 5' regulatory sequence functionally linked to a sequence
encoding a fusion protein OTP/CP4, optionally linked to a 3' regulatory
sequence, wherein said expression cassette functions in plants or plant cells.
|
|
|
AU
759003 B2
- Earliest priority - 24 December 1997
- Filed - 22 December 1998
- Granted - 3 April 2003
- Expected expiry - 22 December 2018
|
Title - Maize H3C4 promoter associated with first rice actin
intron, chimeric gene containing it and transformed plant
Claim 1
A DNA sequence which is a 5' regulatory element allowing the expression of a
heterologous gene in a plant cell from a monocotyledonous plant, characterized
in that it comprises, in the direction of transcription,
a first DNA sequence, which is a functional fragment of the sequence of the
maize H3C4 promoter, and
a second DNA sequence, which is a functional fragment of the sequence of the
first intron of rice actin.
|
Claim 8
A DNA sequence which is a 5' regulatory element allowing the expression of a
heterologous gene in a plant cell from a monocotyledonous plant, characterized
in that it comprises the DNA sequence represented by te
sequence idenntifier No.3
(SEQ
ID NO: 3) or a sequence homologous to the said sequence.
|
|
Aventis CropScience SA (FR)
(now
Bayer
CropScience)
|
|
Remarks
|
Related patents were granted in Europe
(EP
1042491 B1) and China (CN 98813783). A patent
application in Canada (CA 2315677) is still pending. Both the
European patent and the Canadian patent application are in French with an
abstract in English.
|
|
US
5859331
- Earliest priority - 8 July 1992
- Filed - 7 February 1995
- Granted - 12 January 1999
- Expected expiry - 12 January 2016
|
Title - Modular promoter construct
Claim 1
A modular promoter construct, comprising a promoter which is
active in plant cells and a DNA sequence of at least 30 bases from exon 1 of the
rice actin 1 gene, or derivatives of this modular promoter construct which have
promoter activity, wherein said promoter is not a rice actin 1 gene promoter.
|
Claim 5
A vector comprising a promoter construct which comprises a promoter which is
active in plant cells and a DNA sequence of at least 30 bases from exon 1 of the
rice actin 1 gene, or derivatives of this modular promoter construct which have
promoter activity, wherein said promoter construct is coupled to a gene which is
expressed in a plant cell, and wherein said promoter is not a rice actin 1 gene
promoter.
|
Claim 6
A plant cell which is transformed with a vector comprising a promoter
construct which comprises a promoter which is active in plant cells and a DNA
sequence of at least 30 bases from exon 1 of the rice actin 1 gene, or
derivatives of this modular promoter construct which have promoter activity,
wherein said promoter construct is coupled to a gene which is to be expressed in
a plant cell, and wherein said promoter is not a rice actin 1 gene promoter.
|
Claim 7
A plant or its descendants, regenerated from a plant cell comprising a
promoter construct which comprises a promoter which is active in plant cells and
a DNA sequence of at least 30 bases from exon 1 of the rice actin 1 gene, or
derivatives of this modular promoter construct which have promoter activity,
wherein said promoter construct is coupled to a gene which is to be expressed in
a plant cell, and wherein said promoter is not a rice actin 1 gene promoter.
|
Claim 8
A method for preparing plants having elevated gene expression, said method
comprising transforming a plant cell with a vector comprising a promoter
construct which comprises a promoter which is active in plant cells and a DNA
sequence of at least 30 bases from exon 1 of the rice actin 1 gene, or
derivatives of this modular promoter construct which have promoter activity,
wherein said promoter construct is coupled to a gene which is to be expressed in
a plant cell, and wherein said promoter is not a rice actin 1 gene promoter.
|
|
Max-Planck
Institute
|
|
EP
651812 B1
- Earliest priority - 8 July 1992
- Filed - 7 July 1993
- Granted - 15 March 2000
- Expected expiry - 7 July 2013
|
Title - Modular promoter construct
Claim 1
Modular promoter construct, which has a promoter which is active in plant
cells and a DNA sequence from exon 1 of the rice actin 1 gene, and the alleles
and gene expression-stimulating derivatives of this modular promoter construct,
with the proviso that the promoter is not a promoter of the rice actin 1 gene.
|
|
|
Remarks
|
The European patent was converted in Austria (AT), France (FR), Germany (DE),
Italy (IT), Spain (ES), and the Netherlands (NL). The European patent has
abstract and claims in English and the description of the invention is in
German. The granted Australian patent (AU 687004 B2) and the
Canadian patent (CA 2139846) were both lapsed. Application also
filed in Japan (JP 7508653 T2).
|
Note: Patent information was last updated on 22 May 2006. Search terms: "Rice
actin" in abstract. Patent database: PatentLens in combination with INPADOC.
3. Promoter region derived from the rice Act-2 gene
Cornell Research Foundation and Dekalb
Genetics have jointly filed patent applications on the
regulatory elements from the rice Act-2 gene. The promoter
region is referred to as the region that directs the transcriptional activity of
the coding region. As in the rice Act-1 patent, the intron is deemed essential
for promoter efficacy and, in combination with the promoter, is required for the
efficient expression of a gene.
A PCT application
(WO
2000/070067) was filed containing 120 claims, 15 of them
independent. The application discloses an isolated sequence of the 5' region of
the rice Act-2 gene. The region comprises:
- the promoter sequence of about 743 bp,
- exon 1 of the gene,
- intron 1,
- exon 2, and
- the translation initiation codon.
The claims of the PCT application recite:
- isolated rice Act-2 promoter and intron from the 5' region described above,
- an expression vector comprising the rice Act-2 promoter,
- an expression vector comprising the rice Act-2 intron,
- a fertile transgenic plant stably transformed with the rice Act-2 promoter,
- methods to prepare crossed fertile transgenic plants comprising selected
DNA comprising Act-2 promoter and the intron, and
- methods for expressing an exogenous gene in a plant by transforming the
plant with a construct having the gene of interest linked to a rice Act-2
promoter and a rice Act-2 intron.
Note that claims of PCT applications do not provide enforceable
rights and the claim scope may vary in countries that
ultimately grant a patent.
The granted United States patent have similar claim elements but methods for
preparing crossed fertile transgenic plants were excluded.
Specific patent information
|
Patent number
|
Title, Independent Claims and Summary
of Claims
|
Assignee
|
|
US
6429357
- Earliest priority - 14 May 1999
- Filed - 14 May 1999
- Granted - 6 August 2002
- Expected expiry - 14 May 2019
|
Title - Rice actin 2 promoter and intron and methods for
use thereof
Claim 1
An isolated nucleic acid comprising from 40 to about 743 contiguous
nucleotides of the nucleic acid sequence of
SEQ
ID NO:2.
|
Claim 8
An expression vector comprising an isolated rice actin promoter comprising
the nucleic acid sequence of
SEQ
ID NO:1 or
SEQ
ID NO:2 or a fragment thereof having promoter activity.
|
Claim 15
A fertile transgenic plant stably transformed with a selected nucleic acid
comprising a rice actin promoter, wherein said rice actin promoter comprises the
nucleic acid sequence of
SEQ
ID NO:1 or
SEQ
ID NO:2 or a fragment thereof having promoter activity.
|
Claim 26
A method of expressing an exogenous nucleic acid in a plant comprising the
steps of: (i) preparing a construct comprising said exogenous nucleic acid
operably linked to a rice actin promoter, wherein said rice actin promoter
comprises the nucleic acid sequence of
SEQ
ID NO:1 or
SEQ
ID NO:2 or a fragment of
SEQ
ID NO:2 having promoter activity; (ii) transforming a recipient plant cell
with said construct; and (iii) regenerating a transgenic plant expressing said
exogenous nucleic acid from said recipient cell.
|
|
Dekalb Genetics Corp. and Cornell Research Foundation
Inc.
|
|
EP 1179081
- Earliest priority - 14 May 1999
- Filed - 12 May 2000
- Granted - pending
- Expected expiry - N/A
|
Title - Rice actin 2 promoter and intron and methods for use
thereof
The claims are from the corrsponding PCT application WO
2000/070067.
Claim 1
An isolated rice actin 2 promoter isolatable from the nucleic acid sequence
of
SEQ
ID NO:1.
|
Claim 2
An isolated rice actin 2 promoter isolatable from the nucleic acid sequence
of
SEQ
ID NO:2.
|
Claim 3
An isolated nucleic acid comprising from 40 to about 743 contiguous
nucleotides of the nucleic acid sequence of
SEQ
ID NO:2.
|
Claim 10
An isolated rice actin 2 intron isolatable from the nucleic acid sequence of
SEQ
ID NO:1.
|
Claim 11
An isolated rice actin 2 intron isolatable from the nucleic acid sequence of
SEQ
lD NO:3.
|
Claim 12
An isolated nucleic acid comprising from 40 to about 1763 contiguous
nucleotides of the nucleic acid sequence of
SEQ
lD NO:3.
|
Claim 19
An expression vector comprising an isolated rice actin 2 promoter.
|
Claim 32
An expression vector comprising an isolated rice actin 2 intron.
|
Claim 41
A fertile transgenic plant stably transformed with a selected DNA comprising
an actin 2 promoter.
|
Claim 58
A crossed fertile transgenic plant prepared according to the method
comprising the steps of:
(i) obtaining a fertile transgenic plant comprising a selected DNA comprising an
actin 2 promoter;
(ii) crossing said fertile transgenic plant with itself or with a second plant
lacking said selected DNA to prepare the seed of a crossed fertile transgenic
plant comprising said selected DNA; and
(iii) planting said seed to obtain a crossed fertile transgenic plant.
|
Claim 74
A crossed fertile transgenic plant prepared according to the method
comprising:
(i) obtaining a fertile transgenic plant comprising a selected DNA comprising an
actin 2 intron;
(ii) crossing said fertile transgenic plant with itself or with a second plant
lacking said selected DNA to prepare seed of a crossed fertile transgenic plant
comprising said selected DNA; and
(iii) planting said seed to obtain a crossed fertile transgenic plant comprising
said selected DNA.
|
Claim 91
A method of expressing an exogenous gene in a plant comprising the steps of:
(i) preparing a construct comprising said exogenous gene operably linked to an
actin 2 promoter;
(ii) transforming a recipient plant cell with said construct; and
(iii) regenerating a transgenic plant expressing said exogenous gene from said
recipient cell.
|
Claim 98
A method of expressing an exogenous gene in a plant comprising the steps of:
(i) preparing a construct comprising an actin 2 intron and an exogenous gene;
(ii) transforming a recipient plant cell with said construct; and
(iii) regenerating a transgenic plant expressing said exogenous gene from said
recipient cell.
|
Claim 107
A method of plant breeding comprising the steps of:
(i) obtaining a transgenic plant comprising a selected DNA comprising an actin 2
promoter; and
(ii) crossing said transgenic plant with itself or a second plant.
|
Claim 114
A method of plant breeding comprising the steps of
(i) obtaining a transgenic plant comprising a selected DNA comprising an actin 2
intron; and
(ii) crossing said transgenic plant with itself or a second plant.
|
|
CA 2372859
- Earliest priority - 14 May 1999
- Filed - 12 May 2000
- Granted - pending
- Expected expiry - N/A
|
Title - Rice actin 2 promoter and intron and methods for use
thereof
The claims are the same as EP 1179081.
|
|
Remarks
|
The patent application in Australia ( AU 57231/00) lapsed.
|
Note: Patent information was last updated on 23 May 2006. Search terms: "Rice
actin" in abstract. Patent database: PatentLens and esp@cenet in combination
with INPADOC.
Other plant actin promoters
Rice actin promoters have been shown to be strong in directing gene
expression in graminaceous monocotyledonous plants. However, their function in
certain non-graminaceous monocots are variable suggesting that the activity of
actin promoters may be limited to closely related species. Therefore, Researches
were also carried out to identify actin promoters from other plant species. So
far, patents have been issued and patent applications have been filed on actin
promoters from the plant species including banana, melon, maize and
Arabidopsis.
Queensland University of Technology filed patent applications in the United
States and Australia on an actin promoter from banana and its use.
Exelixis Plant Sciences, Inc. obtained patents in United States and
Australia on a melon actin promoter.
Monsanto owns two United States patents and one Australian patent claiming
Arabidopsis actin promoters and their use as part of the invention. In addition,
Monsanto also filed patent applications on actin regulatory elements from rice
and maize in the United States and Europe.
Exelixis Plant Sciences, Inc. patent family
|
Patent/Application No
|
Title, Independent Claims and Summary of Claims
|
Assignee
|
|
US
6642438
- Earliest priority - 19 Mar 1999
- Filed - 17 Mar 2000
- Granted - 04 Nov 2003
- Expected expiry - 17 Mar 2020
|
Title - Melon promoters for expression of transgenes in
plants
Claim 1
An isolated nucleic acid molecule comprising a promoter, wherein the promoter
comprises the nucleic acid sequence presented as
SEQ
ID NO:4.
|
Claim 2
An isolated nucleic acid molecule comprising a promoter, wherein the promoter
consists of a portion of the nucleic acid sequence presented as
SEQ
ID NO:4 that, when operably linked to a protein-encoding polynucleotide
sequence, directs expression of the protein in a plant cell.
|
The claims are directed to a promoter isolated from melon (Cucumis melo) and
its use including:
- An isolated nucleic acid molecule comprising a promoter
- A method for producing a transgenic plant with constitutive expression of
protein encoding polynucleotide sequence under the control of the promoter
(Claim 7).
|
Exelixis Plant Sciences, Inc.
|
|
AU
782602 B2
- Earliest priority - 19 Mar 1999
- Filed - 17 Mar 2000
- Granted - 04 Nov 2003
- Expected expiry - 17 Mar 2020
|
Title - Banana and melon promoters for expression of transgenes in plants
The independent claims of this Australian patent are the same as that in
US
6642438.
|
|
Remarks
|
Patent applications were also filed in Europe (EP 1165755
A1), Canada (CA 2365259) and Japan (JP
2002539779 T2).
|
Note: Patent information was last updated on 9 November 2006. Search terms:
"plant
actin promoter" in abstract. Patent database: PatentLens in combination with
INPADOC.
Monsanto patent family on Arabidopsis actin promoters
|
Patent/Application No
|
Title, Independent Claims and Summary of Claims
|
Assignee
|
|
US
6462258
- Earliest priority - 16 Dec 1999
- Filed - 15 Dec 2000
- Granted - 15 Dec 2000
- Expected expiry - 2 Mar 2021 (adjusted)
|
Title - Plant expression constructs
|
Claim 1
A DNA construct comprising: a first expression cassette, a second expression
cassette, and a third expression cassette, wherein said first expression
cassette in operable linkage comprises
(i) a Figwort mosaic virus 35S promoter;
(ii) a heterologous 5' untranslated leader;
(iii) a chloroplast transit peptide DNA sequence;
(iv) a DNA sequence encoding a glyphosate tolerant EPSPS; and
(v) a transcriptional terminator; and
said second expression cassette comprising in operable linkage
(a) an Arabidopsis Elongation factor 1α promoter with homologous EF1α
intron;
(b) a chloroplast transit peptide DNA sequence;
(d) a DNA sequence encoding a glyphosate tolerant EPSPS; and
(e) a transcriptional terminator; and
said third expression cassette comprising in operable linkage
(I) an Arabidopsis Act2 promoter with homologous Act2 intron;
(II) a chloroplast transit peptide DNA sequence;
(III) a DNA sequence encoding a glyphosate tolerant EPSPS; and
(IV) a transcriptional terminator.
|
|
MONSANTO TECHNOLOGY LLC
|
|
US
6919495
- Earliest priority - 16 Dec 1999
- Filed - 01 May 2003
- Granted - 19 Jul 2005
- Expected expiry - 14 Jun 2023 (adjusted)
|
Title - Chimeric cauliflower mosaic virus 35S-arabidopsis
actin 8 promoters and methods of using them
|
Claim 1
A DNA construct comprising a chimeric promoter DNA sequence comprising
SEQ
ID NO:29; a structural DNA sequence; and a 3' non-translated region that
functions in plants to cause the addition of polyadenylated nucleotides to the
3' end of the RNA sequence; wherein the structural DNA sequence is operably
linked to the chimeric promoter and the 3' non-translated region, and the
chimeric promoter DNA sequence is heterologous with respect to the structural
DNA sequence.
|
|
Claim 7
A DNA construct comprising a chimeric promoter DNA sequence comprising
SEQ
ID NO:29; an aroA:CP4 structural DNA sequence; and a 3' non-translated
region that functions in plants to cause the addition of polyadenylated
nucleotides to the 3' end of the RNA sequence; wherein the structural DNA
sequence is operably linked to the chimeric promoter and the 3' non-translated
region, and the chimeric promoter DNA sequence is heterologous with respect to
the structural DNA sequence.
|
|
Claim 8
A method of expressing a structural DNA sequence in a plant, the method
comprising:
(1) providing a DNA construct comprising a promoter that is functional in
a plant cell, the promoter comprising
SEQ
ID NO:29; a structural DNA sequence; and a 3' non-translated region that
functions to cause the addition of polyadenylated nucleotides to the 3' end of
the RNA sequence; wherein the structural DNA sequence is operably linked to the
promoter and the 3' non-translated region, and the promoter is heterologous with
respect to the structural DNA sequence;
(2) introducing the DNA construct into a plant cell;
(3) regenerating the plant cell to produce the plant and
(4) screening seedlings to identify plants that express the structural
DNA sequence.
|
|
|
AU
782697 B2
- Earliest priority - 16 Dec 1999 (US)
- Filed - 12 Dec 2000
- Granted - 18 Aug 2005
- Expected expiry - 12 Dec 2020
|
Title - DNA constructs for expression of heterologous
polypeptides in plants
|
Claim 1
A hybrid promoter DNA sequence comprising at least one cis element
from a caulimovirus promoter operably linked to at least one cis
element from an Arabidopsis promoter selected from an Act2 promoter,
Act8 promoter or EF1α promoter.
|
|
|
Remarks
|
Related patent applications were also filed in Europe (EP 1632576
A2), Canada (CA 2394984), China (CN
1434868), Japan (JP 2003516753), Brazil (BR
200016460) and Indonesia (ID 28665).
|
Note: Patent information was last updated on 9 November 2006. Search terms:
"actin" in title. Patent database: PatentLens in combination with INPADOC.
Monsanto patent family on on actin regulatory elements from rice and maize
|
Patent/Application No
|
Title, Independent Claims and Summary of Claims
|
Assignee
|
|
US
2006/162010 A1
- Earliest priority - 25 Sep 2003
- Filed - 24 Sep 2004
- Granted - pending
- Expected expiry - N/A
|
Title - Actin regulatory elements for use in plants
Claim 1
An isolated polynucleotide molecule having gene regulatory activity and
comprising a polynucleotide sequence selected from the group consisting of SEQ
ID NO: 1-7.
|
Claim 4
A DNA construct comprising an isolated polynucleotide molecule having gene
regulatory activity and comprising a polynucleotide sequence selected from the
group consisting of SEQ ID NO: 1-7, wherein said isolated polynucleotide
molecule is operably linked to a transcribable polynucleotide molecule.
|
Claim 15
A method of inhibiting weed growth in a field of transgenic glyphosate
tolerant crop plants comprising: i) planting the transgenic plants transformed
with an expression cassette comprising (a) an isolated polynucleotide molecule
having gene regulatory activity and comprising a polynucleotide sequence
selected from the group consisting of SEQ ID NO: 1-7 and operably linked to a
DNA molecule encoding a glyphosate tolerance gene and ii) applying glyphosate to
the field at an application rate that inhibits the growth of weeds, wherein the
growth and yield of the transgenic crop plant is not substantially affected by
the glyphosate application.
|
|
MONSANTO TECHNOLOGY LLC
|
|
Remarks
|
Related European patent application (EP 1664311 A2) was also
filed. Its PCT application is
WO
2005/030968.
|
|
Note: Patent information was last updated on 9 November 2006. Search terms:
"plant actin promoter" in abstract. Patent database: PatentLens in combination
with INPADOC.
Maize alcohol dehydrogenase promoter (Adh)
Scientific aspects
Some plants, e.g., rice, have the ability to synthesize proteins that allow
them to thrive under transient anaerobic conditions. One of those proteins is
alcohol dehydrogenase, which is involved in ethanolic fermentation.
In maize, there are two proteins identified: alcohol dehydrogenase I (ADH-I)
and II (ADH-II), of which ADH-I is the most important because its gene promoter
has been widely used. In the 5' untranslated regions of the maize ADH genes,
anaerobic regulatory elements and an intron are important for driving gene
expression in monocots. The regulatory elements responsible for the anaerobic
response of the genes are within a 247 bp segment immediately upstream of the
CAP site, more specifically between positions -140 and -99 of the maize Adh-1
promoter. Within this 40 bp segment, there are two essential regions, each of
around 15 bp, required for expression under low oxygen conditions.
The whole Adh-1 promoter has been used in cereals such as rice, oat and
barley and in dicots such as tobacco to drive the expression of genes of
interest, providing very low levels of expression in the dicot crop. The Adh-I
promoter in conjunction with the intron has proved to be much superior for
transformation of some cereals such as rice.
Only two regions of the regulatory transcription region of the maize ADH I
gene is currently used to drive gene expression in transformed plants, mainly
monocots:
- the anaerobic regulatory elements (AREs) of the Adh-1
promoter, and
- the first intron of the maize Adh-1 gene.
The two elements are the basis of a recombinant promoter construct called
pEmu. pEmu is formed in the 5' to 3' direction
by:
- a truncated Adh-1 promoter having multiple copies of
ARE,
-
enhancer elements from the octopine synthase
(OCS) gene of A. tumefaciens,
- a TATA box from a promoter expressed in plants, and
- the maize Adh-1 intron 1.
The promoter construct has been tested in transient assays in various monocot
crops, driving a very high level of gene expression compared to the 35S CaMV
promoter. It has also been used for the development of transgenic rice and
sugarcane. However, in some stable transformations, the response has been poor
and the recovery of the transgenic plants difficult. Problems have been
attributed to the presence of duplicated OCS enhancer elements.
IP issues
The Commonwealth Scientific and Industrial Research Organization
(CSIRO) of Australia and the companies Lubrizol
Enterprises and Mycogen Plant Science (U.S.) have
jointly filed and been granted patents related to the Adh promoter and
"regulatory elements". Actual assignment of the patent rights varies
among jurisdictions according to Inpadoc. The table shows the entity identifed
by Inpadoc as owning the patents granted in Australia, Canada, Europe and the
United States. These may now be controlled by Dow.
Approximate scope of protection
The patents do not refer to the maize Adh-1
promoter as such. They cover some elements
derived from the promoter regions of
maize aldolase gene, maize Adh-1 and Adh-2
genes. The patents are in general directed to two different aspects:
- the ARE regions of the promoters of maize ADH genes and maize aldolase gene,
and
- the chimeric pEmu having ARE regions and an intron
1. Patents directed to the ARE enhancer regions
- Patents directed to these regions were granted in the U.S., Canada, and
Europe. The enhancer regions are part of a recombinant promoter molecule that
has, from 5' to 3' direction, at least:
- a promoter expressible in plants, which is basically reduced to a TATA box,
and
- a structural gene under the control of the above mentioned elements
- The claims of the European patent encompass the DNA sequences of the ARE
regions of:
- maize adh-1 and adh-2 genes, and
- sequences that are at least 66% homologous to the mentioned genes
- The Canadian patent also claims a method for the expression of a structural
gene in a plant under conditions of low oxygen.
2. Patents directed to a chimeric promoter known as pEmu
- The patents on a recombinant promoter for enhancing-expression of structural
genes in monocot plant cells have been granted in the
U.S., Europe and Australia. The minimum
elements of the promoter in direction 5' to 3', claimed in all three
jurisdictions, are:
- several ARE enhancer elements
- a TATA box
- a transcription start site
- an intron and
- a structural gene;
- In addition, the independent claims of the U.S. and the
Australian patents comprise enhancer elements derived from the
ocs promoter region.
- Although the examples provided in the specification refer to the ARE regions
and intron 1 derived from the maize aldolase and maize adh genes, the ARE
regions and the intron of the recombinant promoter claimed in the patents in all
three jurisdictions are not limited to the elements of the
maize adh genes. Thus, the use of other ARE enhancer
elements and introns might be protected by the claims of these
patents. The prosecution histories of the patents might shed light on this
aspect.
The patent information, a summary of the independent claims
and the actual independent claims of each patent are presented
in the following tables.
Patents on the ARE of maize ADH and maize aldolase promoters
| Patent number |
Title, Independent Claims and
Summary of Claims
|
Assignee
|
|
US
5001060
- Earliest priority - 6 February 1987
- Filed - 15 June 1987
- Granted - 19 March 1991
- Expected expiry - 18 March 2008
|
Title - Plant Anaerobic regulatory element
Claim 1
A recombinant DNA molecule comprising:
(a) an anaerobic regulatory element;
(b) a plant-expressible promoter located 3' to said anaerobic regulatory
element, and
(c) a plant-expressible structural gene located 3' to said plant-expressible
promoter such that said structural gene is placed under the regulatory control
of said promoter and said anaerobic regulatory element wherein said structural
gene is not in nature under the regulatory control of said anaerobic regulatory
element.
|
- A recombinant DNA molecule comprising from 5' to 3' direction an ARE, a
plant- expressible promoter and a plant-expressible structural gene.
|
Lubrizol Enterprises Inc. &
CSIRO
|
|
CA 1338858
- Earliest priority - 6 February 1987
- Filed - 5 February 1988
- Granted - 21 January 1997
- Expected expiry - 20 January 2014
|
Title - Plant Anaerobic regulatory element
Claim 1
A recombinant DNA molecule comprising: (a) an anaerobic regulatory element;
(b) a plant-expressible promoter located 3' to said anaerobic regulatory
element, and (c) a plant-expressible structural gene located 3' to said
plant-expressible promoter such that said structural gene is placed under the
regulatory control of said promoter and said anaerobic regulatory element
wherein said structural gene is not in nature under the regulatory control of
said anaerobic regulatory element.
|
|
Claim 25
A method for selective expression of a plant-expressible structural gene under
anaerobic conditions in plant tissue which comprises the steps of:
(i) constructing a recombinant DNA moleculae which comprises (a) an anaerobic
regulatory element; (b) a plant-expressible promoter located 3' to said
anaerobic regulatory element, and (c) a plant-expressible structural gene
located 3' to said plant-expressible promoter such that said structural gene is
placed under the regulatory control of said promoter and said anaerobic
regulatory element.
(ii) transforming said plant tissue with said recombinant DNA molecule, and
(iii) placing said transformed plant cell under anaerobic conditions so that
said plant-expressible structural gene is expressed.
|
- A recombinant DNA molecule the same as described in the US patent
US
5001060.
- A method for the anaerobic expression of a plant-expressible structural
gene. The method comprises transforming a plant tissue with a recombinant DNA
molecule as described and placing the tissue under anaerobic conditions to
trigger expression.
|
Mycogen Plant Science Inc. &
CSIRO
|
|
EP
278658 B1
- Earliest priority - 6 February 1987
- Filed - 2 February 1988
- Granted - 10 January 1996
- Expected expiry - 1 February 2008
|
Title - Use of a plant anaerobic regulatory element
|
Claim 1
A method for selective expression of a plant-expressible structural gene under
anaerobic conditions in plant tissue, which method comprises using as an
anaerobic regulatory element a recombinant DNA molecule comprising a sequence
selected from:
1) 5' -GCTGGTTTCT-3'
2) 5' -CGTGGTTTGCTTGCC-3',
or a sequence having about 66% or greater homology thereto
3) 5' -CGAGCCTTTCTTCCC-3'
4) 5' -CTGCCTCCCTGGTTTCT-3', and
5) 5'-CTGCAGCCCCGGTTTCG-3',
or a sequence having about 66% or greater homology thereto,
a plant-expressible promoter being located 3' to said anaerobic regulatory
element, and a plant-expressible structural gene being located 3' to said
plant-expressible promoter such that said structural gene is placed under the
regulatory control of said promoter and said anaerobic regulatory element.
|
- A method for the anaerobic expression of a plant-expressible structural gene
using a recombinant DNA molecule as described in the Canadian patent. The ARE
sequences are selected from maize aldolase gene, regions I and II of the ARE of
maize adh-1 and adh-2. Also, sequences with at least 66% of homology to the
mentioned ones are covered.
|
Lubrizol Enterprises Inc. &
CSIRO
|
| Remarks |
The granted Japanese patent JP 8826414 B2 assigned to Lubrizol Enterprises
& CSIRO was not analyzed. The European patent has lapsed in Belgium (BE),
France (FR), Gemany (DE), Netherlands (NL), Spain (ES), Sweden (SE) and UK.
An application is pending in South Africa (ZA 8800320 A).
|
|
Patents on the recombinant promoter pEMU
|
Patent number
|
Title, Summary of Claims and Independent
Claims
|
Assignee
|
|
EP
459643 B1
- Earliest priority - 19 May 1990
- Filed - 9 May 1991
- Granted - 16 August 2000
- Expected expiry - 8 May 2011
|
Title - A recombinant promoter for gene expression in monocotyledonous plants
Claim 1
A recombinant promoter molecule for enhancing expression of a
plant-expressible structural gene in a monocot plant cell comprising: (a) a
plurality of ARE enhancer elements (b) a truncated, plant expressible promoter
providing a TATA box region necessary to initiate transcription positioned 3' to
said plurality of enhancer elements; and (c) a nucleotide sequence naturally
found as an intron positioned between the transcription start site and the
translation start site in a plant-expressible gene; whereby a plant-expressible
structural gene placed 3' to said recombinant promoter molecule is expressed in
said monocot plant cell under regulatory control of said recombinant promoter
molecule.
|
- A recombinant promoter molecule comprising from 5' to 3' a plurality of ARE
enhancer elements, a promoter providing a TATA box, and an intron. A
plant-expressible gene is located 3' to and under the control of the mentioned
recombinant molecule. The gene is expressed in a monocot plant cell.
|
Mycogen Plant Science Inc. & CSIRO
|
|
US
5290924
- Earliest priority - 19 May 1990
- Filed - 21 April 1993
- Granted - 1 March 1994
- Expected expiry - 20 April 2013
|
Title - Recombinant promoter for gene expression in
monocotyledonous plants
Claim 1
A recombinant promoter molecule, useful for enhancing expression of a
plant-expressible structural gene in a monocot plant cell, said promoter
molecule comprising: (a) a plurality of enhancer elements selected from the
group consisting of only ARE elements, only OCS elements, and combinations of
ARE and OCS elements; (b) a truncated, plant expressible promoter, providing a
TATA box region and a transcription start site, said promoter selected from the
group consisting of Δ35S and ΔADH positioned 3' to said plurality of enhancer
elements wherein said truncated promoter excludes the presence of enhancer
sequences and wherein said truncated promoter is recombined with said plurality
of enhancer elements positioned 5' to said truncated promoter; and (c) a maize
Adh1 intron positioned 3' to said transcription start site whereby a
plant-expressible structural gene, placed 3' to said recombinant promoter
molecule, is expressed in said monocot plant cell under regulatory control of
said recombinant promoter molecule.
|
- A recombinant promoter molecule for expression in monocots similar to the
one claimed in the European patent. The enhancer elements are selected from ARE,
OCS, and a combination of both. There is also a plant-expressible termination
signal 3' to the gene of interest.
|
CSIRO
|
CA 2042831
- Earliest priority - 19 May 1990
- Filed - 17 May 1991
- Granted - 25 July 2000
- Expected expiry - 24 July 2017
|
Title - Recombinant promoter for gene expression in
monocotyledonous plants
Claim 1
A recombinant promoter molecule for enhancing expression of a
plant-expressible structural gene in a monocot plant cell comprising: (a) a
plurality of enhancer elements selected from the group consisting of ARE and OCS
elements; (b) a truncated, plant expressible promoter providing a TATA box
region necessary to initiate transcription positioned 3' to said plurality of
enhancer elements; and (c) a nucleotide sequence naturally found as an intron
positioned between the transcription start site and the translation start site
in a plant-expressible gene; whereby a plant-expressible structural gene placed
3' to said recombinant promoter molecule is expressed in said monocot plant cell
under regulatory control of said recombinant promoter molecule.
|
- A recombinant promoter molecule for enhancing the expression of a gene of
interest in monocots comprising several ARE and OCS elements, a promoter having
a TATA box, an intron. A structural gene is located 3' to the recombinant
promoter.
|
Lubrizol Enterprises Inc. &
CSIRO
|
|
Remarks
|
The granted Australian patent (AU 643521 B2) has lapsed. The
European patent registered in Denmark (DK), France (FR), Germany (DE), Italy
(IT), and Spain (ES).
A patent was also granted in Japan (JP 3325589 B2) and an
application in China (CN 1063506) was withdrawn.
|
|
Note: Patent information on this page was last updated on 13 March 2006.
Synthetic promoters
Scientific aspects
As mentioned in the introductory information about promoters, a set of
minimum elements are required for an active eukaryotic promoter. Among those
elements are the TATA box, the transcription start site or CAP site and the
CCAAT consensus sequence, which is required for accurate transcription.
From the sequences of these elements in diverse organisms, it is possible to
synthesize consensus sequences that may work across different organisms and are
not necessarily derived from a particular organism.
The group of patents under this section are directed to promoters whose parts
are synthesized as consensus sequences of the promoter elements found in nature.
IP issues
Two families of patents directed to synthetic promoters and promoter elements
have been filed by Pioneer Hi-Bred international Inc. The first
patent family includes two granted patents in the U.S. and
Australia and a couple of European
applications and one Canadian application. The second patent
family consists of patent applications filed in Australia, United States,
Europe, Canada and a PCT application. The two patent families are unrelated.
1. The first patent family
Definition of promoter and some of its elements
In the first patent family, promoters are defined in functional and
structural terms. They are described as the transcription control units that
contain the signals for RNA polymerase to begin transcription so that protein
synthesis can occur. Promoters are located in the 5' flanking or upstream region
of the transcribed gene. The most common motifs present in promoters are:
- the TATA element, which is the site where the TATA-binding protein (TBP)
binds. This protein is part of a complex of polypeptides that recruit RNA
polymerase II to begin transcription;
- the transcription start site; and
- the CCAAT consensus sequence.
A core promoter or minimal promoter
contains the TATA box and the transcription start
site. This core promoter may or may not have detectable activity in the
absence of sequence(s) that enhance this activity or confer tissue- specific
activity.
Other elements of promoter regions include:
- INR, sequences near the transcription start site of some genes that provide
an alternate site for binding factors to activate transcription;
- enhancers; and
- upstream elements.
The enhancers are not classified as upstream elements by the
inventors. The upstream elements disclosed, unlike enhancers,
are position and orientation dependent, interact with specific
binding factors and are less common. The upstream elements also may be exchanged
with other elements while maintaining their characteristic control over gene
expression. In contrast, enhancers can increase the efficiency
of transcription regardless of their distance and
orientation to the transcription start site.
Approximate scope of protection
The synthetic promoters that are the subject of the
Pioneer Hi-Bred's inventions contain:
- a TATA motif;
- a GC-rich region (at least 64% GC); and
- a transcription start site.
According to the inventors, the GC-rich region located between the TATA motif
and the transcription start site in plant promoters acts as a very strong
inducer of constitutive expression. It increases transcriptional activation
efficiency. Plant-expressible promoters contain a region of about 40% GC, while
a 64% or greater GC content is characteristic of animal promoters. The maize
ubiquitin 1 gene (Ubi-1) core promoter, which produces high levels of
activity in monocots, has a GC content more similar to animal promoters (64%).
The GC content of promoters covered by the claims is
at least 64%.
Both the U.S. and the Australian patents
claim a synthetic promoter as described. Expression cassettes
containing a structural gene linked to the promoter and a poly(A) signal are
also part of the claimed inventions.
In addition, both patents claim:
- A synthetic upstream element comprising at least 3 octopine
synthase (OCS) binding motifs (TGACG) with an intervening
sequence. This motif has been identified from several opine synthase genes,
i.e., octopine, nopaline, mannopine, and from other genes such as
histone genes.
- Expression cassettes containing the synthetic upstream element linked to the
synthetic promoter.
- Expression cassettes where the synthetic upstream element is linked to a
promoter which does not necessarily have the structural elements described
above.
The U.S. patent also claims a promoter construct having
a core promoter and the upstream activating region
(UAR) of the Ubi-1 gene. An expression
cassette containing these elements, a structural gene linked to the promoter and
a poly(A) signal is also claimed.
A summary of the independent claims of the granted patents
and some bibliographic data are shown in the following table.
| Patent number |
Title, Independent Claims and
Summary of Claims
|
Assignee
|
|
US
6072050
- Earliest priority - 24 February 1998
- Filed - 24 February 1998
- Granted - 6 June 2000
- Expected expiry - 24 February 2018
|
Title - Synthetic promoters
|
Claim 1
A synthetic DNA promoter sequence functional in a plant cell, said promoter
sequence comprising:
a TATA motif,
a transcription start site, and
a region between said TATA motif and said start site that is at least 64%
GC-rich;
wherein said region is not a region between a TATA
motif and a transcription start site of native maize
ubiquitin promoter, and
wherein said promoter sequence is set forth in
SEQ
ID NO:10.
|
|
Claim 2
A synthetic DNA promoter sequence functional in a plant cell, said promoter
sequence comprising:
a TATA motif,
a transcription start site, and
a region between said TATA motif and said start site that is at least 64%
GC-rich;
wherein said region is not a region between a TATA motif and a
transcription start site of native maize ubiquitin promoter, and
wherein said promoter sequence is set forth in
SEQ
ID NO:1.
|
|
Claim 3
An expression cassette comprising
a synthetic promoter comprising:
a TATA motif,
a transcription start site and
a "region" between said TATA motif and said start site that is at
least 64% GC rich,
a structural gene operatively linked to said promoter,
and
a transcription end site polyadenylation signal;
wherein said "region" is not a region
between a TATA motif and a transcription start site of native maize ubiquitin
promoter, and
wherein sequence of said promoter is set forth in
SEQ
ID NO:1.
|
|
Claim 4
An expression cassette comprising
a synthetic promoter comprising:
a TATA motif,
a transcription start site and
a region between said TATA motif and said start site that is at least
64% GC rich,
a structural gene operatively linked to said promoter,
and
a transcription end site polyadenylation signal;
wherein said region is not a region between a TATA
motif and a transcription start site of native maize ubiquitin promoter,
and
wherein sequence of said promoter is set forth in
SEQ
ID NO:10.
|
|
Claim 5
An expression cassette comprising
a synthetic promoter comprising:
a TATA motif,
a transcription start site and
a region between said TATA motif and said start site that is at least
64% GC rich,
a structural gene operatively linked to said promoter,
a transcription end site polyadenylation signal, and
an upstream element operatively linked to said promoter so that
transcription is enhanced;
wherein said region is not a region between a TATA
motif and a transcription start site of native maize ubiquitin promoter;
and
wherein sequence of said upstream element is set forth in
SEQ
ID NO:2.
|
| Claim 7
A synthetic upstream element having a sequence set forth in
SEQ
ID NO:2.
|
|
Claim 8
An expression cassette comprising:
a promoter sequence;
a structural gene operatively linked to said promoter sequence;
a polyadenylation signal; and
a synthetic upstream element comprising
SEQ
ID NO:2 operatively linked to said promoter so that expression is enhanced.
|
| Claim 13
An isolated nucleotide sequence comprising a DNA enhancer
sequence comprising the nucleotide sequence set forth in SEQ ID
No: 5.
|
| Claim 14
A nucleotide sequence comprising a promoter construct,
said construct comprising in operable linkage
a core promoter sequence and
a Ubi-1 UAR,
wherein said Ubi-1 UAR is a maize Ubi UAR comprising
the sequence set forth in
SEQ
ID No:13.
|
|
Claim 15
An expression cassette comprising in operable linkage:
a core promoter sequence,
a Ubi UAR operably linked upstream to said core promoter to form a synthetic
promoter construct,
a nucleotide sequence of interest operably linked to said synthetic
promoter, and
a polyadenylation signal;
wherein said Ubi-1 UAR comprises the sequence set forth
in
SEQ
ID No:13.
|
The claims are directed to:
- Synthetic promoters functional in plants comprising a TATA motif, a region
at least 64% GC-rich, and a transcription start site. The DNA sequence of the
promoters are either as defined in SEQ ID NO:10 or SEQ ID NO:1 (claims 1 and 2).
The sequences of both are fairly similar, except that one includes G/C
transversions.
- Expression cassettes comprising these synthetic promoters and a
structural gene linked to the promoter and a poly (A) signal
(see claims 3-5).
- A synthetic upstream element (SEQ ID NO:2).
- An expression cassette as described having in addition the above mentioned
upstream element linked to a promoter (claim 8). To be covered by the claim
language, the promoter in this case does not necessarily have the structural
elements (TATA motif, GC-richness, or transcription start site) described above,
but must enhance the transcription process.
- An isolated DNA sequence comprising a DNA enhancer sequence comprising the
nucleotide sequence TGACG (SEQ ID No: 5).
- A nucleotide sequence comprising a promoter construct comprising a core
promoter sequence and a maize Ubi-1 UAR.
- An expression cassette comprising a Ubi-1 UAR linked upstream to a
core promoter, a nucleotide sequence of interest linked to the promoter and a
poly (A) signal.
According to the specification, 64% or greater GC content from
TATA to start of transcription is
generally characteristic of animal promoters. Thus, claims 1-5 are potentially
targeting the use of animal promoters in plant systems.
'Having' in claim 7 is ambiguous; would an infringing sequence contain only
this, or this and other elements?
The isolated nucleotide sequence as an enhancer in claim 13 can have
nucleotides in addition to the TGACG motif. Using bigger
sequences containing TGACG could still infrige the claim.
The Ubi-1 UAR (upstream activating region) claimed in claims
14 and 15 comprises the sequence disclosed in SEQ ID No:13. The Ubi-1 UAR can be
used in both directions and in multiple copies. However, the Ubi-1 UAR sequence
disclosed here is 813 bp and use only portions of the sequence would not be
covered by these claims.
|
Pioneer Hi-Bred International Inc.
|
|
US
6555673
- Earliest priority - 24 February 1998
- Filed - 21 April 2000
- Granted - 29 April 2003
- Expected expiry - 21 April 2020
|
Title - Synthetic promoters
Claim 1
A promoter construct comprising:
a) a synthetic core promoter functional in a plant cell,
wherein said synthetic core promoter has a sequence
comprising
a TATA motif,
a transcription start site, and
a region between said TATA motif and said start site that is at
least 64% GC-rich,
wherein said sequence is set forth in
SEQ
ID NO:10;
b) a synthetic upstream element of
SEQ
ID NO:2 operatively linked to said synthetic core promoter so that control
of transcription from said synthetic core promoter is enhanced;
and
c) at least one upstream activating region.
|
Claim 14
A promoter construct comprising:
a) a synthetic core promoter functional in a plant cell,
wherein said synthetic core promoter has a sequence
comprising
a TATA motif, a transcription start site, and
a region between said TATA motif and said start site that is at
least 64% GC-rich,
wherein said sequence is set forth in
SEQ
ID NO:1;
b) a synthetic upstream element of
SEQ
ID NO:2 operatively linked to said core promoter so that control of
transcription from said core promoter is enhanced; and
c) at least one upstream activating region.
|
Claim 15
A promoter construct comprising:
a) a synthetic core promoter functional in a plant cell,
wherein said synthetic core promoter has a sequence
comprising
a TATA motif,
a transcription start site, and
a region between said TATA motif and said start site that is at
least 64% GC-rich,
wherein said sequence is set forth
inSEQ
ID NO:1;
b) a synthetic upstream element of
SEQ
ID NO:2 operatively linked to said core promoter so that control of
transcription from said core promoter is enhanced; and
c) at least one upstream activating region, wherein at least one of said
upstream activating region(s) is selected from the group
consisting of Ubi-1 UAR and CaMV 35S UAR.
|
Claim 16
A promoter construct comprising:
a) a synthetic core promoter functional in a plant cell,
wherein said synthetic core promoter has a sequence
comprising
a TATA motif,
a transcription start site, and
a region between said TATA motif and said start site that is at
least 64% GC-rich,
wherein said sequence is set forth in
SEQ
ID NO:10;
b) a synthetic upstream element of
SEQ
ID NO:2 operatively linked to said core promoter so that control of
transcription from said core promoter is enhanced; and
c) at least one upstream activating region, wherein at
least one of said upstream activating region(s) is selected from the group
consisting of Ubi-1 UAR and CaMV 35S UAR.
|
Claim 17
A promoter construct comprising:
a) a synthetic core promoter functional in a plant cell,
wherein said synthetic core promoter has a sequence comprising
a TATA motif,
a transcription start site, and
a region between said TATA motif and said start site that is at
least 64% GC-rich,
and wherein said core promoter is a synthetic core promoter
comprising the sequence set forth in
SEQ
ID NO:1; and
b) at least two upstream activating regions, wherein at
least one of said upstream activating regions is selected from the group
consisting of Ubi-1 UAR and CaMV 35S UAR.
|
Claim 18
A promoter construct comprising:
a) a synthetic core promoter functional in a plant cell,
wherein said synthetic core promoter has a sequence
comprising
a TATA motif,
a transcription start site, and
a region between said TATA motif and said start site that is at
least 64% GC-rich,
and wherein said core promoter is a synthetic core promoter
comprising the sequence set forth in
SEQ
ID NO:10; and
b) at least two upstream activating regions, wherein at
least one of said upstream activating regions is selected from the group
consisting of Ubi-1 UAR and CaMV 35S UAR.
|
Claim 19
A promoter construct comprising the sequence set forth in
SEQ
ID NO:12.
|
Claim 20
A promoter construct comprising the sequence set forth in
SEQ
ID NO:15.
|
Claim 21
A promoter construct comprising the sequence set forth in
SEQ
ID NO:16.
|
Claim 22
A promoter construct comprising the sequence set forth in
SEQ
ID NO:17.
|
Claim 23
A promoter construct comprising the sequence set forth in
SEQ
ID NO:18.
|
Claim 24
A promoter construct comprising a sequence having at least
95% sequence identity to the portion of
SEQ
ID NO:12, which is not a CaMV 35S UAR sequence wherein
control of transcription from said promoter is enhanced in comparison to
transcription from a promoter consisting of the synthetic core promoter sequence
set forth in
SEQ
ID NO:1.
|
Claim 25
A promoter construct comprising a sequence having at least
95% sequence identity to the portion of
SEQ
ID NO:15 which is not a Ubi-1 UAR sequence, wherein control
of transcription from said promoter is enhanced in comparison to transcription
from a promoter consisting of the synthetic core promoter sequence set forth in
SEQ
ID NO:1.
|
Claim 26
A promoter construct comprising a sequence having at least
95% sequence identity to the portion of
SEQ
ID NO:16 which is not a Ubi-1 UAR sequence, wherein control of transcription
from said promoter is enhanced in comparison to transcription from a promoter
consisting of the synthetic core promoter sequence set forth in
SEQ
ID NO:1.
|
Claim 27
A promoter construct comprising a sequence having at least
95% sequence identity to the portion of
SEQ
ID NO:17 which is not a Ubi-1 UAR sequence, wherein control of transcription
from said promoter is enhanced in comparison to transcription from a promoter
consisting of the synthetic core promoter sequence set forth in
SEQ
ID NO:1.
|
Claim 28
A promoter construct comprising a sequence having at least
95% sequence identity to the portion of
SEQ
ID NO:18 which is not a Ubi-1 UAR sequence, wherein control of transcription
from said promoter is enhanced in comparison to transcription from a promoter
consisting of the synthetic core promoter sequence set forth in
SEQ
ID NO:1.
|
This patent is a Continuation of
US
6072050.
The claims are drawn to:
1. Promoter constructs comprise:
- a synthetic core promoter which has a sequence (SEQ ID NO:10 or SEQ ID NO:1)
comprising a TATA motif, a transcription start site, and a region between the
TATA motif and the start site that is at least 64% GC-rich
- a synthetic upstream element (SEQ ID NO:2) linked to synthetic core promoter
so that control of transcription from the synthetic core promoter is enhanced
- at least one upstream activating region
2. Promoter constructs comprising any of the following sequences:
SCP1 promoter, SEQ ID NO:12; UCP1 promoter,
SEQ ID NO:15; UCP2 promoter, SEQ ID NO:16;
UCP3 promoter, SEQ ID NO:17 and UCP4
promoter, SEQ ID NO:18). (see claims 19-23)
3. A promoter construct comprising a sequence having at least 95% sequence
identity to the portion of the SCP1 promoter, which is not a
CaMV 35S UAR sequence. This promoter can direct higher transcription than a
promoter consisting of the synthetic core promoter sequence (SEQ ID NO:1) (see
claim 24).
4. A promoter construct comprising a sequence having at least 95% sequence
identity to the portion of one of the promoters: UCP1 promoter,
UCP2 promoter, UCP3 promoter, and
UCP4 promoter, which is not a Ubi-1 UAR sequence.These
promoters can direct higher transcription than a promoter consisting of the
synthetic core promoter sequence (SEQ ID NO:1) (see claims 25-28).
This patent claims promoter constructs comprising all the claim elements in
US
6072050. In addition, promoters with at least 95% identity
to the portion of promoters SCP1 (35S UAR linked to core promoter
of SEQ ID NO: 1), UCP1 (one copy of Ubi1
UAR linked with the core promoter), UCP2
(2 copies of Ubi1 UAR linked with the core
promoter), UCP3 (3 copies of Ubi1 UAR
linked with the core promoter) and UCP4
(4 copies of Ubi1 UAR linked with the core
promoter) are also covered.
According to the specification, upstream
activating region (UAR) is typically a position or
orientation dependent element that primarily directs tissue, cell type, or
regulated expression. The UAR in claims 1 and 14 is not
restricted to Ubi-1 UAR and CaMV 35S UAR and therefore could be from any
organism.
|
|
AU
751402 B2
- Earliest priority - 24 february 1998
- Filed - 23 February 1999
- Granted - 15 August 2002
- Expected expiry -23 February 2019
|
Title - Synthetic promoters
Claim 1
A synthetic DNA plant promoter sequence, said sequence
comprising:
a TATA motif;
a transcription start site;
a region between said TATA motif and said start site that is at least about
64% GC-rich,
an upstream element; and
one or more upstream activating regions;
wherein said promoter sequence comprises a sequence selected
from the group consisting of:
a) a nucleotide sequeuce set forth in SEQ ID NO; 12,
b) a nucleotide sequence set forth in SEQ ID NO: 15,
c) a nucleotide sequence set forth in SEQ ID NO: 16,
d) a nucleotide sequence set forth in SEQ ID NO: 17, and
e) a nucleotide sequence set forth in SEQ ID NO: 18.
|
Claim 2
An expression cassette comprising:
a synthetic promoter comprising
a TATA motif;
a transcription start site and a region there between that is at least
about 64% GC rich;
an upstream element; and
one or more upstream activating regions;
a structural gene operatively linked to said promoter; and
a transcription end site polyadenylation signal, wherein said promoter
sequence comprises a sequence selected from the group
consisting of:
a) a. nucleotide sequence set forth in SEQ ID NO: 12,
b) a nucleotide sequence set forth in SEQ ID NO: 15,
c) a nucleotide sequence set forth in SEQ ID NO: 16,
d) a nucleotide sequence set Forth in SEQ ID NO: 17, and
e) a nucleatide sequence set forth in SEQ ID NO: 18.
|
Claim 10
An expression cassette comprising:
a synthetic core promoter comprising the sequence set forth
in SEQ ID NO:1 or SEQ ID NO:10,
a synthetic upstream element comprising the sequence set
forth in SEQ ID NO:2, and
an upstream activating region comprising the sequence set
forth in SEQ ID NO:12 or 13,
a structural gene operatively linked to said promoter; and
a transcription end site polyadenylation signal.
|
Claim 11
A DNA sequence comprising a promoter construct, said
construct
comprising in operable linkage:
a core synthetic promoter sequence comprising
a TATA motif,
a transcription start site, and
a region between said TATA motif and said start site that is at least
64% GC-rich,
wherein said region is not a region between a TATA
motif and a transcription start site of native maize ubiquitin promoter;
a heterologous upstream element; and
a heterologous upstream activating region operably linked to said core
synthetic promoter; wherein said upstream activating region is selected from the
group consisting of CaMV 35S UAR and Ubi-1 UAR.
|
Claim 19
A method for controlling the level of expression of a transgenic nucleotide
sequence in a dicotyledonous plant cell, said method comprising
transforming said plant cell with an expression cassette
comprising
a synthetic promoter comprising
a TATA motif;
a transcription start site and a region there between that is at least
about 64% GC-rich;
an upstream element; and
one or more upstream activating regions;
a structural gene operatively linked to said promoter; and
a transcription end site polyadenylation signal,
wherein said promoter sequence comprises a sequence
selected from the group consisting of:
a) a nucleotide sequeuce set forth in SEQ ID NO; 12,
b) a nucleotide sequence set forth in SEQ ID NO: 15,
c) a nucleotide sequence set forth in SEQ ID NO: 16,
d) a nucleotide sequence set forth in SEQ ID NO: 17, and
e) a nucleotide sequence set forth in SEQ ID NO: 18.
|
Claim 34
A method for controlling the level of expression of a transgenic nucleotide
sequence in a dicotyledonous plant cell, substantially as hereinbefore described
with reference to any one of the examples.
|
The Australian patent claims cover all the claim elements for the synthetic
promoters, the upstream element and the UARs as claimed in
US
6555673. Addition to these, it also claims for a method for
controlling the level of expression of a transgenic nucleotide sequence in
dicotyledonous plant cells comprising transforming a plant cell with an
expression cassette comprising a synthetic promoter comprising the above claim
elements.
|
|
Remarks
|
Related applications are pending in Europe (EP 1056875) and
Canada (CA 2314598). A patent was also granted in New Zealand
(NZ 506182).
|
|
AU
729929 B2
- Earliest priority - 11 June 1996
- Filed - 10 June 1997
- Granted - 15 February 2001
- Expected expiry - 10 June 2017
|
Title - A synthetic plant core promoter and upstream
regulatory element
|
Claim 1
A synthetic DNA plant promoter sequence, said sequence comprising:
a TATA motif,
a transcription start site, and
a region between said TATA motif and said start site that is at least 64%
GC-rich.
|
|
Claim 3
An expression cassette comprising
a synthetic promoter comprising
a TATA motif,
a transcription start site and
a region there between that is at least 65% GC rich,
a structural gene operatively linked to said promoter, and
a transcription end site polyadenylation signal.
|
| Claim 17
A synthetic upstream element having a sequence of SEQ ID NO:2.
|
|
Claim 18
An expression cassette comprising:
a promoter sequence;
a structural gene operatively linked to said promoter sequence;
a polyadenylation signal; and
a synthetic upstream element comprising SEQ ID NO:2 operatively linked to said
promoter sequence so that expression is enhanced.
|
|
Claim 23
A synthetic DNA plant promoter sequence functional in a plant cell, said
sequence comprising:
a TATA motif,
a transcription start site, and
a region between said TATA motif and said start site that is at least 64%
GC-rich; wherein said promoter sequence is less than 1000 bp.
|
The claims are related to:
- A synthetic promoter comprising a TATA motif, a region at least 64% GC-rich,
and a transcription start site.
- An expression cassette comprising a synthetic promoter and a structural gene
linked to the promoter and a poly (A) signal.
- A synthetic upstream element as claimed in the related U.S. patent
(US
6072050).
- An expression cassette as described having in addition the the above
upstream element linked to the promoter. The element enhances the transcription
process.
- A synthetic promoter functional in plant cells. The elements are the same as
described for the former synthetic promoter but this one is limited to maximum 1
kb in length.
The claims for the synthetic promoter in this Australian patent is very broad
as comparing to the related US and Canadian patents.
|
|
EP
914454 B1
- Earliest priority - 11 June 1996
- Filed - 10 June 1997
- Granted - 9 March 2005
- Expected expiry - 9 June 2017
|
Title - A synthetic plant core promoter and upstream
regulatory element
Claim 1
A synthetic upstream element having a sequence of SEQ ID NO:2.
|
Claim 2
An expression cassette comprising:
a promoter;
a structural gene operatively linked to said promoter sequence;
a polyadenylation signal; and
a synthetic upstream element comprising SEQ ID NO:2 operatively linked to said
promoter so that expression is enhanced.
|
Claim 6
A nucleic acid vector comprising
(i) a promoter operatively linked to a structural gene, wherein the promoter is
a synthetic DNA plant promoter sequence which comprises:
a TATA motif;
a transcription start site; and
a region between the TATA motif and the start site that is at least 64% GC-rich;
and
(ii) an upstream element operatively linked to said promoter comprising the
sequence of SEQ ID NO:2.
|
|
|
CA 2257719
- Earliest priority - 11 June 1996
- Filed - 10 June 1997
- Granted - 31 May 2005
- Expected expiry - 9 June 2017
|
Title - A synthetic plant core promoter and upstream
regulatory element
|
Claim 1
A synthetic DNA promoter functional in plant cells, said promoter comprising:
a TATA motif,
a transcription start site, and
a region between said TATA motif and said start site that is at least 64%
GC-rich, wherein the sequence of said promoter is set forth in SEQ ID NO:10.
|
Claim 2
A synthetic DNA promoter functional in plant cells, said promoter comprising:
a TATA motif,
a transcription start site, and
a region between said TATA motif and said start site that is at least 64%
GC-rich, wherein the sequence of said promoter is set forth in SEQ ID NO:1.
|
Claim 3
An expression cassette comprising:
a synthetic promoter functional in plant cells, wherein the sequence of said
promoter is set forth in SEQ ID NO:1;
a structural gene operatively linked to said promoter; and
a transcription end site polyadenylation signal.
|
Claim 4
An expression cassette comprising:
a synthetic promoter functional in plant cells, wherein the sequence of said
promoter is set forth in SEQ ID NO:1;
a structural gene operatively linked to said promoter; and
a transcription end site polyadenylation signal.
|
Claim 16
A synthetic upstream element having a sequence of SEQ ID NO:2.
|
| Claim 18
An expression cassette comprising:
a promoter sequence;
a structural gene operatively linked to said promoter sequence;
a polyadenylation signal; and
a synthetic upstream element comprising SEQ ID NO:2 operatively linked to said
promoter so that expression is enhanced.
|
|
|
Remarks
|
Application also filed in Brazil (BR 9710690 A)
|
2. Second Patent Family
This patent family has patent applications filed in the United States,
Europe, Australia and Canada. However, the applications in the US, Europe and
Canada have lapsed. For the purpose of providing the disclosed art as prior
art, only brief information about this patent family is provide here, but it
should be noted that the application may still be pending in Australia, and
continuations may still be pending elsewhere.
Some bibliographic data is outlined in the following table:
| Applicant |
Title |
Publication No. |
Publication Date |
|
Pioneer Hi-Bred International Inc.
|
Novel plant promoters and methods of use
|
US
2001/47092 A1
|
29 November 2001
|
|
|
WO
01/53476 A2
|
26 July 2001
|
|
|
EP 1252304
|
30 October 2002
|
|
|
AU 34507/01
|
31 July 2001
|
|
|
CA 2390753
|
26 July 2001
|
What's disclosed in this patent family
The patent applications are directed to nucleotide sequences of plant
promoters comprising synthetic multimeric promoter elements regions
(SMPERs). The different sequences of the SMPERs contain the following
promoter elements:
- the dehydration or drought responsive element (DRE) of
Arabidopsis, which is part of a stress-responsive gene promoter;
- the abscisic acid responsive element (ABRE) of maize, which
acts in embryo development;
- the activating sequence (AS-1) of the Cauliflower mosaic
virus (CaMV), which mediates both salicylic acid- and auxin-inducible
transcriptional activation;
- the DNA-binding domain of the rice transcription factor
GT-2; and
- the proliferating cell nuclear antigen (PCNA) IIA gene
promoter element of rice, which confers meristematic tissue-specific expression.
The SMPERs can be linked to any other promoters, native or synthetic. The
above-mentioned promoter elements function as transcription factor binding
sites. Plants or parts of plants containing one SMPER linked to an encoding
sequence and enhancing the expression of the coding sequence are claimed.
Methods for selecting promoter elements and for creating SMPERs active in a
tissue of interest are also claimed.
Note: Patent information on this page was last updated on 29 March 2006.
Chapter 3
Inducible promoters
Summary
As their name says, the activity of these promoters is induced by the
presence or absence of biotic or abiotic factors. Inducible promoters are a very
powerful tool in genetic engineering because the expression of genes operably
linked to them can be turned on or off at certain stages of development of an
organism or in a particular tissue.
This section presents a general view of patents directed to promoters whose
activity is triggered by either chemical or physical factors. The patents
discussed in this section are about general promoter systems and their uses and
do not encompass particular promoters. There are virtually hundreds of inducible
promoters that vary according to the organism source and cells or tissues where
they regulate gene transcription. Thus, this section only discusses patents
drawn to inducible promoter systems that are regulated by particular chemical or
physical factors.
The analyzed patents on inducible promoters are grouped as:
-
Chemically-regulated promoters,
including promoters whose transcriptional activity is regulated by the presence
or absence of alcohol, tetracycline, steroids, metal and other compounds.
-
Physically-regulated promoters,
including promoters whose transcriptional activity is regulated by the presence
or absence of light and low or high temperatures.
Chemically-regulated promoters
Summary
The activity of this class of promoters is modulated by chemical compounds
that either turn off or turn on gene transcription. As prerequisites, the
chemicals influencing promoter activity typically
- should not be naturally present in the organism where expression of the
transgene is sought;
- should not be toxic;
- should affect only the expression of the gene of interest;
- should be easy to apply or removal; and
- should induce a clearly detectable expression pattern of either high or very
low gene expression
for their optimal use as modulators of gene expression.
Preferably, chemically-regulated promoters should be derived from organisms
distant in evolution to the organisms where its action is required. For example,
promoters to be used in plants are mostly derived from organisms such as yeast,
E. coli, Drosophila or mammals.
This section presents an analysis of patents disclosing some of the main
chemically-inducible promoters actually used to modulate expression in plants
and animals. The promoter types analyzed are grouped as follows:
-
Alcohol-regulated: Syngenta has
several patents and patent applications in Europe and Australia directed to a
transcriptional system containing the alcohol dehydrogenase I (alcA) gene
promoter and the transactivator protein AlcR. Different agricultural
alcohol-based formulations are used to control the expression of a gene of
interest linked to the alcA promoter.
-
Tetracycline-regulated: Yale
University and BASF AG have several patents and patent
applications filed in the United States, Europe, Australia and Canada covering
aspects of tetracycline-responsive promoter systems, which can function
either to activate or repress gene expression system in the presence of
tetracycline. Some of the elements of the systems include a tetracycline
repressor protein (TetR), a tetracycline operator sequence (tetO) and a
tetracycline transactivator fusion protein (tTA), which is the fusion of TetR
and a herpes simplex virus protein 16 (VP16) activation sequence. Eukaryotic
cells transformed with the promoter systems including animal cells are claimed.
-
Steroid-regulated: Numerous patent and patent
applications are directed to steroid-responsive promoters for the modulation of
gene expression in plant and animal cells. Analysis on patents on this type of
promoters includes:
-
McGill University patents on promoters based on the rat
glucocorticoid receptor (GR);
-
Rockefeller University patents on promoters based on the
human estrogen receptor (ER);
-
Syngenta and Pioneer Hi-Bred patents
directed to promoters based on ecdysone receptors derived from different moth
species; and
- a group of patents filed by different entities covering promoters from the
steroid/retinoid/thyroid receptor superfamily.
-
Metal-regulated: Promoters derived from
metallothionein (proteins that bind and sequester metal ions) genes from yeast,
mouse and human are the subject matter of several United States patents granted
to Genentech, University Patents Inc. and
The University of California (Berkeley). DNA
constructs having metal-regulated promoters and eukaryotic cells transformed
with them are claimed.
-
Pathogenesis-related (PR) proteins are induced in
plants in the presence of particular exogenous chemicals in addition to being
induced by pathogen infection. Salicylic acid, ethylene and benzothiadiazole
(BTH) are some of the inducers of PR proteins. Promoters derived from
Arabidopsis and maize PR genes are the subject matter of patents granted to
Novartis and Pioneer Hi-Bred in the United
States, Australia and Europe.
Alcohol-regulated promoters
Scientific aspects
The promoter system in its natural environment
In a bacterial positive-mode operon or positive
operon the genes are off by default and are turned on by
activators. The interaction between a regulatory
protein and a regulatory region on the DNA to which it
binds turns transcription on.
The ethanol utilization pathway of the filamentous fungus Aspergillus
nidulans is a well-characterized positive operon system. It controls the
cellular response to ethanol and other related chemicals. The first enzyme in
the pathway of ethanol utilization is alcohol dehydrogenase I
(Adh-I) encoded by the alcA gene. The
transcriptional activator protein AlcR binds target sequences
within the alcA gene promoter in the presence of
ethanol, ethyl methyl ketone or other alcohols/ketones. These compounds act as
inducers of the gene expression.
A promoter system transferred into plants
One system transferred into plants provides the alcR
gene under the control of a strong constitutive promtoer such as
CaMV 35S, and a modfied alcA promoter
linked to a gene of interest or target gene. The modified or
chimeric alcA promoter (referred to as the target
promoter) comprises the regulatory sequence of the
alcA promoter which is bound by AlcR and a core promoter region (a TATA
box and a transcription start site) of a plant-expressible gene promoter. In the
presence of an inducer (e.g., an alcohol or ketone), the transactivator protein
AlcR binds to the specific sequences of the modified alcA promoter and
the target gene is expressed.
Among the advantages of the system are:
- the lack of an equivalent alcR gene in plants, which rules out
interference with plant endogenous Adh activity,
- the chemical inducers are relatively simple molecules with low toxicity, and
- under normal growth conditions of a plant, the levels of natural inducers,
(e.g., ethanol) that are produced when plants are waterlogged or under
artificial anoxia are very low and do not induce alcA expression.
The alcohol-inducible system can be used for:
- production of high levels of recombinant proteins, especially those that
interfere with growth or biomass production,
- providing an inducible male sterility system, and
- providing inducible expression of resistance genes.
Nevertheless, there are still some difficulties in using the system for
agricultural purposes. Specifically, the means to introduce the inducer, e.g.,
by spraying or drenching, and its volatility, impact the effectiveness of the
system for field work.
IP issues
Syngenta has patents related to alcohol-inducible expression
systems for plants. Its patents were granted in United States and Europe.
Approximate scope of protection
The only patent grant is in Europe. The patent claims:
- a chemically-inducible plant gene expression cassette that contains:
- a promoter linked to an AlcR encoding gene, and
- an inducible promoter linked to a gene of interest.
- a method for controlling plant gene expression by using the mentioned
expression cassette, and
- a chimeric chemically-inducible promoter containing:
- regulatory sequences of the alcA gene promoter,
and
- a transcription initiation sequence from a plant-expressible gene promoter.
Note that the inducible promoter of the expression cassette is
not restricted to any particular promoter and may be other than
alcA (claim 1). The promoter is activatable by the regulatory protein
(AlcR protein) in the presence of an effective inducer. The inducible promoter
may be derived from aldehyde dehydrogenase gene or other alcohol dehydrogenase
genes that are involved in the ethanol utilization pathway.
The transcription initiation sequence of the chimeric chemically-inducible
promoter refers to the CAAT box, TATA box and surrounding sequences that define
the transcription start point for the structural gene.
Specific Patent Information
|
Patent Number
|
Title, Summary of Claims and Independent Claims
|
Assignee
|
|
EP
637339 B1
- Earliest priority -13 April 1992
- Filed - 13 April 1993
- Granted - 31 October 2001
- Expected expiry - 13 April 2013
|
Title - DNA constructs and plants incorporating them
Claim 1
A chemically-inducible plant gene expression cassette
comprising
a first promoter operatively linked to the alcR regulator sequence
obtainable from Aspergillus nidulans which encodes the AlcR regulator
protein, and
an inducible promoter operatively linked to a target gene, the inducible
promoter being activated by the regulator protein in the presence of an
effective exogenous inducer whereby application of the inducer causes expression
of the target gene.
|
Claim 11
A method for controlling plant gene expression comprising
transforming a plant cell with a chemically-inducible plant gene expression
cassette which has
a first promoter operatively linked to the alcR regulator sequence
obtainable from Aspergillus nidulans which encodes the AlcR regulator
protein, and
an inducible promoter operatively linked to a target gene, the inducible
promoter being activated by the regulator protein in the presence of an
effective exogenous inducer whereby application of the inducer causes expression
of the target gene.
|
Claim 12
A chimeric promoter comprising
an upstream region containing a promoter regulatory sequence obtainable from
the alcA gene promoter of Aspergillus nidulans and
a downstream region containing a transcription initiation sequence,
characterised in that said upstream and downstream regions are heterologous, the
promoter is chemically inducible and the transcription initiation sequence is
obtainable from the core promoter region of a promoter which is active in plant
cells.
|
The claims are generally to:
- A chemically-inducible expression cassette comprising a first promoter
linked to the alcR sequence and an inducible promoter linked to a
target gene. The AlcR protein activates the inducible promoter in the presence
of an inducer and causes the expression of the gene.
- A chimeric promoter comprising an upstream region with a
chemically-inducible alcA gene promoter and a downstream region with a
transcription initiation sequence from a core plant-expressible promoter region,
in which the two regions are heterologous.
- A method for controlling plant gene expression by transforming a plant with
an expression cassette as described.
|
Syngenta
Ltd.
|
|
US
6605754
- Earliest priority -13 April 1992
- Filed - 13 March 1998
- Granted 12 August 2003
- Expected expiry - 12 March 2018
|
Title - DNA constructs and plants incorporating them
Claim 1
A plant cell which contains stably integrated into its genome a gene
expression cassette, said gene expression cassette comprising
a first promoter operatively linked to a sequence comprising an alcR coding
sequence from Aspergillus nidulans and which encodes an ALCR regulator
protein, and
an inducible promoter from an ALCR-activatable gene, which gene is the alcA
gene from Aspergillus nidulans, operatively linked to a target gene,
said inducible promoter being activated by the ALCR regulator protein in the
presence of an alcohol and/or ketone inducer, so that application of a
sufficient amount of a suitable inducer causes expression of the target gene.
|
Claim 7
A method for controlling plant gene expression comprising
transforming a plant cell with a chemically-inducible gene expression
cassette which has
a first promoter operatively linked to a sequence comprising an alcR
coding sequence from Aspergillus nidulans, and which encodes an ALCR
regulator protein, and
an inducible promoter from an ALCR-activatable gene, which gene is the
alcA gene from Aspergillus nidulans, operatively linked to a target
gene, said inducible promoter being activated by the ALCR regulator protein in
the presence of an alcohol and/or ketone inducer, so that application of a
sufficient amount of a suitable inducer causes expression of the target gene.
|
Claim 8
A plant cell containing a chimeric promoter operatively linked to a target
gene, said chimeric promoter comprising
an upstream region containing a promoter from an ALCR-activatable alcA gene
from Aspergillus nidulans and a downstream region containing a
transcription initiation sequence, wherein the upstream region and the
downstream region are heterologous and the chimeric promoter is inducible by an
alcohol and/or ketone, so that application of a sufficient amount of a suitable
inducer, in the presence of the ALCR regulator protein encoded by the alcR gene
from Aspergillus nidulans, causes expression of the target gene.
|
The claims of this US patent is genearlly directed to:
- Plant cells containing a chemically-inducible plant gene expression cassette
or chimeric promoter operatively linked to a target gene as described in
EP
637339 B1.
- A method for controlling plant gene expression by transforming a plant cell
with the chemically-inducible gene expression cassette as described so that gene
expression is controlled by an alcohol and/or ketone inducer.
|
|
Remarks
|
The application in Australia (AU 39019/93 A1) has lapsed.
|
Note: Patent information was last updated on 11 May 2006. Search terms:
"promoter" in abstract and "Syngenta" in application. Patent database:
PatentLens in combination with INPADOC.
Tetracycline-regulated promoters
Scientific aspects
Transposons are mobile genetic elements which can insert at random into
plasmids or a bacterial chromosome independently of the host cell recombination
system. Transposons carry genes that confer new phenotypes on the host cell
such as antibiotic resistance.
The Tetracycline resistance operon is carried by the
Escherichia coli transposon (Tn) 10. This operon has a
negative mode of operation. The interaction between a
repressor protein encoded by the
operon,TetR, and a DNA sequence to which it
binds, the tet operator (tetO), represses the activity
of a promoter placed near the operator. In the absence of an inducer,TetR binds
to tetO and prevents transcription. Transcription can be turned on when an
inducer, such as tetracycline, binds to TetR and causes a conformation change
that prevents TetR from remaining bound to the operator. When the operator site
is not bound, the activity of the promoter is restored.
In plants, the system has been adapted in two ways
- as a promoter repressing system, basically using the system as described,
and
- as a promoter activating system, where TetR is modified to activate
gene transcription, instead of inhibiting transcription.
In one system, a CaMV 35S promoter is modified by introducing a tet
operator sequence upstream and downstream of the TATA box. In the
absence of tetracycline, overexpressed TetR binds to
the tet operator and prevents gene expression. In the presence of
tetracycline, TetR no longer binds the operator and gene expression is turned
on.
In tobacco, the expression of the tetracycline-inducible CaMV promoter could
be modulated up to 500-fold. This inducible promoter has also worked in tomato
and potato.
Unfortunately, the system presents some problems. For tetracycline to work as
an inducer, it must be supplied continuously to the medium due in part to the
short-half life of the antibiotic. In addition, TetR must be in high
concentration to be effective as a repressor as it has to compete with at least
forty proteins that assemble around the TATA box. For some plants, such as
Arabidopsis, high concentrations of the repressor are toxic and alter
the photosynthetic physiology of the plant.
Due to the drawbacks of the promoter repressing system, TetR has also been
converted to an activator of gene expression. In one version
of a promoter activating system, TetR is fused to the acidic activation sequence
of the herpes simplex virus protein 16 (VP16), forming a tetracycline
transactivator (tTA) fusion protein which is has the DNA
binding specificity of TetR and the promoter activating function of VP16. In the
absence of tetracycline, tTA binds to tet operator
sequences placed upstream of a TATA box in a target promoter and activates
transcription. When tetracycline is provided, it forms a complex with tTA and
releases the operator, thus, turning off gene transcription.
In contrast to the wild type TetR, tTA does not need to
compete with endogenous transcription factors for binding sites. The system has
worked in tobacco and in Arabidopsis.
Despite the advantages of the Tet activating system, the plants must be in
the presence of tetracycline to turn transcription off. That implies a
continuous supply of the antibiotic. Also the original promoter that contains
the tet operators is prone to silencing over time. Work has been done
to improve the efficacy of the promoter.
IP issues
Yale University and BASF
AG have several patents and patent applications related to
tetracycline-regulated promoter systems. The inventions claimed by these
institutes include the use of a wild-type TetR and fusion proteins formed by a
wild type TetR or a mutated TetR linked to either an activator or an inhibitor
of transcription.
1. Yale University patents
Yale has a United States patent and a European patent directed to an
auto-regulated tetracycline transactivator tTA
placed under the control of a minimal promoter having at least one
tetO sequence.
The inventions
In the United States patent
US
5851796 and the European patent
EP
832254 B1, the production of tTA is regulated by the presence of
tetracycline: in its presence, tTA is not produced. In the absence of
tetracycline, tTA binds the tetO sequence promoting the expression of the
gene(s) under the control of an inducible promoter.
Claims are drawn to:
A polynucleotide:
- encoding a tetracycline transactivator fusion protein and
- operably linked to an inducible minimal promtoer with at least one tetO
sequence
The United States patent describes that a gene that's operably linked to a
promoter sequence is placed under the control of that sequence.
The encoded protein contains:
- a prokaryotic tet repressor (the claim is not limited to only the DNA
binding portion) and a
- eukaryotic transcriptional activator protein (the activator is not limited
to the VP16 activator).
Although the patent describes that different activator sequences can be used
(listing acidic rich domains such as in VP16, proline rich domains of CTF/NF-1,
serine/threonine rich domains of Oct-2, or glutamine rich domains, such as found
in Sp1), the only construct tested is one in which the activator is VP16.
The specification describes that a "eukaryotic transcriptional activator" is
capable of activating transcription in eukaryotes.
Methods for decreasing or enhancing the expression of a heterologous protein
in a eukaryotic cell are also claimed. The methods generally include inserting
two DNA molecules into the cell: One of them is the polynucleotide described
above.; the other encodes a heterologous protein to be regulated by the system
linked to a minimal promoter, which also has at least a tetO sequence. When the
transformed eukaryotic cell is cultivated in the presence of tetracycline or a
tetracycline analogue, the expression of the heterologous protein is inhibited
because the activator tTA does not remain bound to the minimal promoter having
the tetO sequence that drives the expression of the heterologous gene.
Conversely, in the absence of tetracycline, the heterologous protein is
expressed in the eukaryotic cell because the tTA activator remains bound
The specification defines "heterologous" as is a protein that does not
naturally occur in the specific host organism in which it is present.
Kits containing the two types of molecules used in the method are also
claimed. However, the kits have to have:
- at least two container means each containing a different one of the two
molecules;
- the means have to be in close confinement in the kit.
An additional kit claim requires that:
- the mentioned polynucleotide encoding the transactivator fusion protein is
within a eukaryotic cell.
The patent does not describe what a "container means" is or how close "close
confinement" has to be.
The specific patent information of
US
5851796 and
EP
832254 B1 is presented in the following table. For more information
about licencing etc., the Yale
Office of Cooperative Research shoud be contacted.
|
Patent number
|
Title, Independent Claims and
Summary of Claims
|
Assignee
|
|
US
5851796
- Earliest priority - 7 June 1995
- Filed - 7 June 1995
- Granted - 22 December 1998
- Expected expiry - 22 December 2015
|
Title - Autoregulatory tetracycline-regulated system for
inducible gene expression in eucaryotes
Claim 1
A polynucleotide comprising a nucleotide sequence encoding a
tetracycline transactivator fusion protein, said protein
comprising a prokaryotic tet repressor and a eucaryotic
transcriptional activator protein, and said polynucleotide molecule being
operably linked to an inducible minimal promoter, which promoter contains at
least one tet operator sequence.
|
Claim 23
A method to inhibit expression of a heterologous protein in a eucaryotic cell
comprising
(a) obtaining a eucaryotic cell comprising
(i) a first polynucleotide molecule encoding a tetracyline
transactivator fusion protein, said protein comprising a
procaryotic tet repressor and a eucaryotic transcriptional activator protein,
and said polynucleotide molecule being operably linked to an inducible minimal
promoter, which promoter contains at least one tet operator sequence;
(ii) a second polynucleotide molecule encoding the heterologous protein,
said second polynucleotide molecule being operably linked to an inducible
minimal promoter, and said promoter containing at least one tet operator
sequence; and
(b) cultivating the eucaryotic cell in a medium comprising tetracycline or a
tetracycline analogue such that expression of the heterologous protein is
inhibited.
|
Claim 25
A method to enhance the expression of a heterologous protein in a eucaryotic
cell comprising
(a) obtaining a eucaryotic cell comprising
(i) a first polynucleotide molecule encoding a tetracycline
transactivator fusion protein, said protein comprising a
prokaryotic tet repressor and a eucaryotic transcriptional activator protein,
and said polynucleotide molecule being operably linked to an inducible minimal
promoter, which promoter contains at least one tet operator sequence;
(ii) a second polynucleotide molecule encoding the heterologous protein,
said second polynucleotide molecule being operably linked to an inducible
minimal promoter, and said promoter containing at least one tet operator
sequence; and
(b) cultivating the eucaryotic cell in a medium lacking tetracycline or a
tetracycline analogue such that expression of the heterologous protein is
enhanced.
|
Claim 26
A method to activate the expression of a heterologous protein in a eucaryotic
cell comprising
(a) obtaining a eucaryotic cell comprising
(i) a first polynucleotide molecule encoding a tetracycline
transactivator fusion protein, said protein comprising a
prokaryotic tet repressor and a eucaryotic transcriptional activator protein,
and said polynucleotide molecule being operably linked to an inducible minimal
promoter, which promoter contains at least one tet operator sequence;
(ii) a second polynucleotide molecule encoding the heterologous protein,
said second polynucleotide molecule being operably linked to an inducible
minimal promoter, and said promoter containing at least one tet operator
sequence; and
(b) cultivating the eucaryotic cell in a medium lacking tetracycline or a
tetracycline analogue such that expression of the heterologous protein is
activated.
|
Claim 27
A kit comprising a carrier means having in close confinement
therein at least two container means,
wherein a first container means contains a first polynucleotide molecule
encoding a tetracycline transactivator fusion protein, said protein
comprising a procaryotic tet repressor and a eucaryotic
transcriptional activator protein, and said polynucleotide molecule being
operably linked to an inducible minimal promoter, which promoter contains at
least one tet operator sequence; and
a second container means contains a second polynucleotide molecule
encoding said inducible minimal promoter, which promoter
contains at least one tet operator sequence, which tet operator sequence is
strategically positioned for being operably linked to a heterologous
polynucleotide sequence encoding a polypeptide.
|
Claim 28
A kit comprising a carrier means having in close confinement
therein at least two container means,
wherein a first container means contains a eucaryotic cell transfected
with a first polynucleotide molecule encoding a tetracycline transactivator
fusion protein, said protein comprising a procaryotic tet
repressor and a eucaryotic transcriptional activator protein, and said
polynucleotide molecule being operably linked to an inducible minimal promoter,
which promoter contains at least one tet operator sequence; and
a second container means contains a second polynucleotide molecule
comprising an inducible minimal promoter, which promoter
contains at least one tet operator sequence, which tet operator sequence is
strategically positioned for being operably linked to a heterologous
polynucleotide sequence encoding a heterologous polypeptide.
|
|
Yale University
|
|
EP
832254 B1
- Earliest priority - 7 June 1995
- Filed - 7 June 1996
- Granted - 29 March 2006
- Expected expiry - 7 June 2015
|
Title - Autoregulatory tetracycline-regulated system for
inducible gene expression in eucaryotes
|
Claim 1
A composition of matter comprising a polynucleotide molecule
encoding a tetracycline transactivator fusion protein, said protein
comprising a prokaryotic tet repressor and a eucaryotic
transcriptional activator protein, and said polynucleotide molecule being
operably linked to an inducible minimal promoter, which promoter contains at
least one tet operator sequence, wherein the open reading frame of the
polynucleotide molecule encoding the tetracycline transactivator fusion protein
is modified at its 5' end to provide an optimal context for translational
initiation.
|
|
Claim 11
A method to decrease or shut off expression of a heterologous protein
comprising
(a) transforming a eucaryotic cell with
(i) a first polynucleotide molecule encoding a tetracyline
transactivator fusion protein, said protein comprising a
prokaryotic tet repressor and a eucaryotic transciptional activator protein, and
said polynucleotide molecule being operably linked to an incudicble minimal
promoter, which promoter contains at least one tet operator sequence, wherein
the open reading frame of the polynucleotide molecule encoding the tetracycline
transactivator fusion protein is modified at its 5' end to provide an optimal
context for translational initiation;
(ii) a second polynucleotide molecule encoding the heterologous
protein, said protein being operably linked to an inducible minimal promoter,
and said promoter containing at least one tet operator sequence; and
(b) cultivating the eucaryotic cell in a medium comprising tetracycline or a
tetracycline analogue.
|
|
Claim 13
A method to activate or enhance the expression of a heterologous protein
comprising
(a) transforming a eucaryotic cell with
(i) a first polynucleotide molecule encoding tetracycline transactivator
fusion protein, said protein comprising a prokaryotic tet repressor and a
eucaryotic transciptional activator protein, and said polynucleotide molecule
being operably linked to an inducible promoter, which promoter contains at least
one tet operator sequence, wherein the open reading frame of the polynucleotide
molecule encoding the tetracycline transactivator fusion protein is modified at
its 5' end to provide an optimal context for translational initiation;
(ii) a second polynucleotide molecule encoding the heterologous
protein, said protein being operably linked to an inducible minimal promoter,
and said promoter containing at least one tet operator sequence; and
(b) cultivating the eucaryotic cell in a medium lacking tetracycline or a
tetracycline analogue.
|
|
Claim 17
A composition of matter consisting essentially of the plasmid pTet-Splice as
represented in Figure 9A, wherein pTet-Splice comprises SEQ ID NO: 2.
|
|
Claim 18
A composition of matter consisting essentially of the plasmid pTet-tTAK as
represented in Figure 10A, wherein pTet-tTAK comprises SEQ ID NO: 3.
|
|
Claim 19
A kit comprising a carrier means having in close confinement
therein at least two container means,
wherein a first container means contains a first polynucleotide molecule
encoding a tetracycline transactivator fusion protein, said protein
comprising a procaryotic tet repressor and a eucaryotic
transcriptional activator protein, and said polynucleotide molecule being
operably linked to an inducible minimal promoter, which promoter contains at
least one tet operator sequence, wherein the open reading frame of the
polynucleotide molecule encoding the tetracycline transactivator fusion protein
is modified at its 5' end to provide an optimal context for translational
initiation; and
a second container means contains a second polynucleotide molecule encoding
said inducible minimal promoter, which promoter contains at least one tet
operator sequence, which tet operator sequence is strategically positioned for
being operably linked to a heterologous polynucleotide sequence encoding a
polypeptide.
|
|
Claim 20
A kit comprising a carrier means having in close confinement
therein at least two container means,
wherein a first container means contains a eucaryotic cell transfected with
a first polynucleotide molecule encoding a tetracycline transactivator fusion
protein, said protein comprising a procaryotic tet repressor
and a eucaryotic transcriptional activator protein, said polynucleotide molecule
being operably linked to an inducible minimal promoter, which promoter contains
at least one tet operator sequence, wherein the open reading frame of the
polynucleotide molecule encoding the tetracycline transactivator fusion protein
is modified at its 5' end to provide an optimal context for translational
initiation; and
a second container means contains a second polynucleotide molecule
comprising an inducible minimal promoter, which promoter
contains at least one tet operator sequence, which tet operator sequence is
strategically positioned for being operably linked to a heterologous
polynucleotide sequence encoding a heterologous polypeptide.
|
The independent claims are similar to the claims of
US
5851796, except for the metioning of the
5' end modification of the open reading frame of the polynucleotide molecule
encoding the tetracycline transactivator fusion protein and the claims for the
composition of matter consisting essentially of the plasmids pTet-Splice and
pTet-tTAK.
|
|
Remarks
|
The application filed in Australia (AU 62745/96) has lapsed.
Related application also filed in Japan (JP 11507539 T2).
|
Note: Patent information on this page was last updated on 15 March 2006.
Search terms: "tetracycline" in abstract and "Yale University" in applicant.
Patent database: PatentLens in combination with INPADOC.
2. BASF AG patents
This company has a large portfolio of granted patents and patent applications
related to tetracycline-regulated promoters. Patents have been granted in
Australia (1), Canada (1) and in the United States (16). Additional applications
have been filed in Australia, Canada, China and Europe. The inventions cover
different versions of Tet-activating systems, transgenic organisms transformed
with tetracycline-regulated promoters including eukaryotes in general, and
plants and animals in particular.
Detailed information on the BASF AG patents is provided in the following
table. For more information, please contact
BASF
AG .
|
Patent Number
|
Title, Independent Claims and Summary of Claims
|
Assignee
|
|
US
5464758
- Earliest priority - 14 June 1993
- Filed - 14 June 1993
- Granted - 7 November 1995
- Expected expiry - 14 June 2013
|
Title - Tight control of gene expression in
eucaryotic cells by tetracycline-responsive promoters
Claim 1
A polynucleotide molecule coding for a transactivator fusion protein
comprising a prokaryotic tet repressor and a eucaryotic transcriptional
activator protein domain.
|
Claim 10
A polynucleotide molecule coding for a protein, wherein said polynucleotide
is operably linked to a minimal promoter and at least one tet operator sequence.
|
Claim 16
A eucaryotic cell transfected with
(a) a first polynucleotide molecule coding for a transactivator fusion
protein comprising a prokaryotic tet repressor and a eucaryotic transcriptional
activator protein domain; and
(b) a second polynucleotide molecule coding for a protein, wherein said
second polynucleotide molecule is operably linked to a minimal promoter and at
least one tet operator sequence.
|
Claim 18
A kit comprising a carrier means having in close confinement
therein at least two container means,
wherein a first container means contains a first polynucleotide molecule
coding for a transactivator fusion protein comprising a
prokaryotic tet repressor and a eucaryotic transcriptional activator protein
domain, and
a second container means contains a second polynucleotide molecule
comprising a minimal promoter operably linked to at least one
tet operator sequence, wherein said minimal promoter is capable of being ligated
to a heterologous gene sequence coding for a polypeptide.
|
Claim 19
A kit comprising a carrier means having in close confinement
therein at least two container means, wherein
a first container means contains a eucaryotic cell transfected with a first
polynucleotide molecule coding for a transactivator fusion protein
comprising a prokaryotic tet repressor and a eucaryotic
transcriptional activator protein domain, and
a second container means contains a second polynucleotide molecule
comprising a minimal promoter operably linked to at least one
tet operator sequence, wherein said minimal promoter is capable of being ligated
to a heterologous gene sequence coding for a polypeptide.
|
|
BASF AG
|
|
US
5650298
- Earliest priority - 14 June 1993
- Filed - 14 June 1994
- Granted - 22 July 1997
- Expected expiry - 22 July 2014
|
Title - Tight control of gene expression in eucaryotic cells
by tetracycline-responsive promoters
Claim 1
An isolated DNA molecule for integrating a polynucleotide sequence encoding a
tetracycline-controllable transactivator (tTA) at a predetermined location in a
second target DNA molecule, the tTA comprising a prokaryotic
Tet repressor operably linked to a polypeptide which directly or indirectly
activates transcription in eucaryotic cells, the DNA molecule
comprising a polynucleotide sequence encoding the tTA flanked
at 5' and 3' ends by additional polynucleotide sequences of sufficient length
for homologous recombination between the DNA molecule and the second target DNA
molecule at a predetermined location.
|
Claim 11
An isolated DNA molecule for integrating a polynucleotide sequence encoding a
tetracycline-controllable transactivator (tTA) and a tTA-responsive promoter
within a predetermined gene of interest in a second target DNA molecule, the DNA
molecule comprising:
a) a first polynucleotide sequence comprising a 5' flanking regulatory
region of the gene of interest, operably linked to:
b) a second polynucleotide sequence encoding a tTA, the tTA comprising a
prokaryotic Tet repressor operably linked to a polypeptide which directly or
indirectly activates transcription in eucaryotic cells; and
c) a third polynucleotide sequence comprising a tTA-responsive promoter,
operably linked to:
d) a fourth polynucleotide sequence comprising at least a portion of a
coding region of the gene of interest;
wherein the first and fourth polynucleotide sequences are of sufficient
length for homologous recombination between the DNA molecule and the gene of
interest in the second target DNA molecule such that expression of the tTA is
controlled by 5' regulatory elements of the gene of interest and expression of
the gene of interest is controlled by the tTA-responsive promoter.
|
Claim 45
A method for producing a host cell having a DNA molecule encoding a
tetracycline-controllable transactivator (tTA) integrated at a predetermined
location in a second target DNA molecule within the cell,
comprising:
a) introducing the DNA molecule of claim 1 into a population of cells
under conditions suitable for homologous recombination between the DNA encoding
the tTA and the second target DNA molecule; and
b) selecting a cell in which the DNA encoding the tTA has integrated at a
predetermined location within the second target DNA molecule.
|
|
|
AU
684524 B2
- Earliest priority - 14 June 1993
- Filed - 14 June 1994
- Granted - 18 December 1997
- Expected expiry - 14 June 2014
|
Title - Tight control of gene expression in eucaryotic cells
by tetracycline-responsive promoters
Claim 1
The same as Claim 1 of
US
5650298.
|
Claim 11
The same as Claim 11 of
US
5650298.
|
Claim 42
A non-human transgenic animal having a transgene comprising
a polynucleotide sequence encoding a tetracycline-controllable transactivator
(tTA), the tTA comprising a prokaryotic Tet repressor operably
linked to a polypeptide which directly or indirectly activates transcription in
eucaryotic cells.
|
| Claim 52
A non-human transgenic animal having a transgene comprising
a polynucleotide sequence encoding a tetracycline-controllable transactivator
(tTA), the tTA comprising a prokaryotic Tet repressor operably
linked to a polypeptide which directly or indirectly activates transcription in
eucaryotic cells, wherein the transgene is integrated by homologous
recombination at a predetermined location within a chromosome within cells of
the animal.
|
| Claim 55
A transgenic animal having a transgene comprising
a polynucleotide sequence encoding a tetracycline-controllable
transactivator (tTA) and
a tTA-responsive promoter,
wherein the transgene is integrated by homologous recombination at a
predetermined location within a gene of interest within cells of the animal such
that expression of the tTA is controlled by 5' regulatory elements of the gene
of interest and expression of the gene of interest is controlled by the
tTA-responsive promoter.
|
|
|
CA 2165162
- Earliest priority - 14 June 1993
- Filed - 14 June 1994
- Granted - 23 May 2000
- Expected expiry - 14 June 2014
|
Title - Tight control of gene expression in eucaryotic cells
by tetracycline-responsive promoters
Claim 1
The same as Claim 1 of
US
5650298.
|
Claim 11
The same as Claim 11 of
US
5650298.
|
Claim 42
Use of tetracycline or a tetracycline analogue for the inhibition of a second
transgene in a transgenic animal, said animal having
a first transgene comprising a polynucleotide sequence
encoding a tetracycline-controllable transactivator (tTA), the tTA comprising a
prokaryotic Tet repressor operably linked to a polypeptide which directly or
indirectly activates transcription in eucaryotic cells; and
the second transgene comprising a gene of interest operably
linked to a tTA-responsive promoter.
|
|
Claim 43
The use of tetracycline or a tetracycline analogue for the inhibition of
transcription of a second transgene in a transgenic animal, said animal having
a polynucleotide sequence encoding a tetracycline-controllable
transactivator (tTA), the tTA comprising a prokaryotic Tet repressor operably
linked to a polypeptide which directly or indirectly activates transcription in
eucaryotic cells, wherein the first transgene is integrated by homologous
rrecombination at a predetermined location within a chromosome within cells of
the animal; and
the second transgene comprising a gene of interest operably
linked to a tTA-responsive promoter.
|
Claim 44
Use of tetracycline or a tetracycline analogue for inhibiting transcription
of the gene of interest in a transgenic animal having a transgene
comprising
a polynucleotide sequence encoding a tetracycline-controllable
transactivator (tTA) and
a tTA-responsive promoter,
wherein the transgene is integrated by homologous rrecombination at a
predetermined location within a gene of interest within cells of the animal such
that expression of the tTA is controlled by 5' regulatory elements of the gene
of interest and expression of the gene of interest is controlled by the
tTA-responsive promoter.
|
Claim 48
A method for producing a non-human transgenic animal
comprising:
a) introducing a DNA molecule encoding the tTA into a fertilized oocyte;
b) implanting the fertilized oocyte in a pseudopregnant foster mother;
and
c) allowing the fertilized oocyte to develop into the non-human
transgenic animal to thereby produce the non-human transgenic animal.
|
|
|
|
Remarks
|
The related European patent application (EP 705334 A2) is
pending.
|
|
|
US
5654168
- Earliest priority - 14 June 1993
- Filed - 15 July 1994
- Granted - 5 August 1997
- Expected expiry - 5 August 2014
|
Title - Tetracycline-inducible transcriptional activator and
tetracycline-regulated transcription unit
Claim 1
An isolated nucleic acid encoding a fusion protein which activates
transcription, the fusion protein comprising a first
polypeptide which binds to a tet operator sequence in the presence of
tetracycline or a tetracycline analogue operatively linked to a second
polypeptide which activates transcription in eukaryotic cells.
|
Claim 26
A kit comprising a carrier means having in close confinement
therein at least two container means comprising:
a) a first container means containing a first nucleic acid encoding a
fusion protein which activates transcription, the fusion protein comprising a
polypeptide which binds to a first class of tet operator sequence in the
presence of tetracycline or a tetracycline analogue operatively linked to a
polypeptide which activates transcription in enkaryotic cells;
and
b) a second container means containing a second nucleic acid comprising a
first cloning site for introduction of a first nucleotide sequence to be
transcribed operatively linked to at least one tet operator sequence of a first
class type.
|
|
|
|
US
5789156
- Earliest priority - 14 June 1993
- Filed - 3 February 1995
- Granted - 4 August 1998
- Expected expiry - 4 August 2015
|
Title - Tetracycline-regulated transcriptional inhibitors
Claim 1
An isolated nucleic acid encoding a fusion protein which inhibits
transcription in eukaryotic cells, the fusion protein
comprising a first polypeptide which binds to tet operator
sequences, operatively linked to a heterologous second polypeptide which
inhibits transcription in eukaryotic cells.
|
Claim 46
A kit comprising a carrier means having in close confinement therein at least
two container means comprising:
a) a first container means containing a first nucleic acid encoding a
fusion protein which inhibits transcription in eukaryotic cells, the fusion
protein comprising a first polypeptide which binds to tet operator sequences
either
(I) in the presence but not the absence of tetracycline or a
tetracycline analogue or
(ii) in the absence but not the presence of tetracycline or a
tetracycline analogue, operatively linked to a heterologous second polypeptide
which inhibits transcription in eukaryotic cells, or a eukaryotic cell line into
which said first nucleic acid has been stably introduced; and
b) a second container means containing a second nucleic acid comprising a
cloning site for introduction of a nucleotide sequence to be transcribed
operatively linked to at least one tet operator sequence.
|
This patent is a Continuation in part of
US
5654168.
|
|
|
US
5589362
- Earliest priority - 14 June 1993
- Filed - 7 June 1995
- Granted - 31 December 1996
- Expected expiry - 31 December 2013
|
Title - Tetracycline-regulated transcriptional modulators
with altered DNA binding specificities
Claim 1
An isolated nucleic acid molecule encoding a fusion protein which regulates
transcription, the fusion protein comprising a Tet repressor
having at least one amino acid mutation that confers on the fusion protein an
ability to bind a class B tet operator sequence having a nucleotide substitution
at position +4 or +6, operatively linked to a polypeptide which regulates
transcription in eukaryotic cells.
|
Claim 13
A method for regulating transcription of a tet operator-linked gene in an
isolated cell, comprising:
introducing into the isolated cell a nucleic acid molecule encoding a
fusion protein which regulates transcription, the fusion protein
comprising a Tet repressor having at least one amino acid
mutation that confers on the fusion protein an ability to bind a class B tet
operator sequence having a nucleotide substitution at position 14 or 16,
operatively linked to a polypeptide which regulates transcription in eukaryotic
cells; and
modulating the concentration of a tetracycline, or analogue thereof, in
contact with the isolated cell.
|
This patent is a Continuation in part of
US
5789156.
|
|
|
US
5814618
- Earliest priority - 14 June 1993
- Filed - 7 June 1995
- Granted - 29 September 1998
- Expected expiry - 29 September 2015
|
Title - Methods for regulating gene expression
Claim 1
A method for regulating expression of a tet operator-linked gene in a cell
of a subject, comprising:
introducing into the cell a nucleic acid molecule encoding a fusion
protein which inhibits transcription in eukaryotic cells, the fusion protein
comprising a first polypeptide which binds to a tet operator
sequence, operatively linked to a heterologous second polypeptide which inhibits
transcription in eukaryotic cells; and
modulating the concentration of a tetracycline, or analogue thereof, in
the subject.
|
Claim 17
A method for regulating expression of a gene in a cell of a subject,
comprising:
obtaining the cell from the subject;
introducing into the cell a first nucleic acid molecule which operatively
links a gene to at least one tet operator sequence;
introducing into the cell a second nucleic acid molecule encoding a
fusion protein which inhibits transcription, the fusion protein
comprising a first polypeptide which binds to a tet operator
sequence, operatively linked to a second polypeptide which inhibits
transcription in eukaryotic cells, to form a modified cell;
administering the modified cell to the subject; and
modulating the concentration of a tetracycline, or analogue thereof, in
the subject.
|
This patent is a Continuation in part of
US
5789156.
|
|
|
US
5888981
- Earliest priority - 14 June 1993
- Filed - 7 June 1995
- Granted - 30 March 1999
- Expected expiry - 30 March 2016
|
Title - Methods for regulating gene expression
Claim 1
A method for regulating expression of a tet operator-linked gene in a cell of
a subject, comprising:
introducing into the cell a nucleic acid molecule encoding a
tetracycline-controllable transactivator (tTA), the tTA comprising a Tet
repressor operably linked to a polypeptide which directly or indirectly
activates transcription in eucaryotic cells; and
modulating the concentration of a tetracycline, or analogue thereof, in
the subject.
|
Claim 10
A method for regulating expression of a gene in a cell of a subject,
comprising:
obtaining the cell from the subject;
introducing into the cell a first nucleic acid molecule which operatively
links a gene to at least one tet operator sequence;
introducing into the cell a second nucleic acid molecule encoding a
tetracycline-controllable transactivator (tTA), the tTA
comprising a Tet repressor operably linked to a polypeptide
which directly or indirectly activates transcription in eucaryotic cells, to
form a modified cell;
administering the modified cell to the subject; and
modulating the concentration of a tetracycline, or analogue thereof, in
the subject.
|
This patent is a Continuation in part of
US
5650298.
|
|
|
US
6004941
- Earliest priority - 14 June 1993
- Filed - 7 June 1995
- Granted - 21 December 1999
- Expected expiry - 21 December 2016
|
Title - Methods for regulating gene expression
Claim 1
A method for regulating expression of a tet operator-linked gene in a cell of
a subject, comprising:
introducing into the cell a nucleic acid molecule encoding a fusion
protein which activates transcription, the fusion protein
comprising a first polypeptide which binds to a tet operator
sequence in the presence of tetracycline or a tetracycline analogue operatively
linked to a second polypeptide which activates transcription in eukaryotic
cells; and
modulating the concentration of a tetracycline, or analogue thereof, in
the subject, such that expression of a tet operator-linked gene in a cell of the
subject is regulated.
|
Claim 16
A method for regulating expression of a gene in a cell of a subject,
comprising:
obtaining the cell from the subject;
introducing into the cell a first nucleic acid molecule which operatively
links a gene to at least one tet operator sequence;
introducing into the cell a second nucleic acid molecule encoding a
fusion protein which activates transcription, the fusion protein
comprising a first polypeptide which binds to a tet operator
sequence in the presence of tetracycline or a tetracycline analogue operatively
linked to a second polypeptide which activates transcription in eukaryotic
cells, to form a modified cell;
administering the modified cell to the subject; and
modulating the concentration of a tetracycline, or analogue thereof, in
the subject such that expression of the gene which is operatively linked to at
least one tet operator sequence is regulated in a cell of the subject.
|
Claim 28
An isolated recombinant vector for coordinate, bidirectional transcription of
a first and a second nucleotide sequence, the vector comprising
a nucleotide sequence comprising in a 5' to 3' direction:
a) a first cloning site for introduction of a first nucleotide sequence
to be transcribed, operatively linked to
b) at least one tet operator sequence, operatively linked to
c) a second cloning site for introduction of a second nucleotide sequence
transcribed,
the vector further comprising additional regulatory
sequences such that the vector is sufficient for use in eukaryotic cells,
wherein transcription of a first and second nucleotide sequence introduced into
the vector proceeds in opposite directions relative to the at least one tet
operator sequence.
|
Claim 35
An isolated nucleic acid composition comprising at least one
recombinant vector for independent regulation of transcription of a first and a
second nucleotide sequence, the nucleic acid composition comprising nucleotide
sequences comprising:
a) a first cloning site for introduction of a first nucleotide sequence
to be transcribed, operatively linked to at least one let operator sequence of a
first class type; and
b) a second cloning site for introduction of a second nucleotide sequence
to be transcribed, operatively linked to at least one let operator sequence of a
second class type.
|
This patent is a Continuation in part of
US
5789156.
|
|
|
US
5859310
- Earliest priority - 14 June 1993
- Filed - 7 June 1995
- Granted - 12 January 1999
- Expected expiry - 12 January 2016
|
Title - Mice transgenic for a tetracycline-controlled transcriptional
activator
Claim 1
A transgenic mouse having a transgene integrated into the genome of the mouse
and also having a tet operator-linked gene in the genome of the mouse, wherein:
the transgene comprises a transcriptional regulatory element
functional in cells of the mouse operatively linked to a polynucleotide sequence
encoding a tetracycline-controllable transactivator fusion protein (tTA),
said fusion protein comprises a Tet repressor operably
linked to a polypeptide which directly or indirectly activates transcription of
said tet operator-linked gene in eucaryotic cells,
said tet operator-linked gene confers a detectable and functional
phenotype on the mouse when expressed in cells of the mouse,
said transgene is expressed in cells of the mouse at a level sufficient
to produce amounts of said fission protein that are sufficient to activate
transcription of the tet operator-linked gene; and
in the absence of tetracycline or a tetracycline analogue in the mouse,
said fusion protein binds to the tet operator-linked gene and activates
transcription of the tet operator linked gene such that the tet operator-linked
gene is expressed at a level sufficient to confer the detectable and functional
phenotype on the mouse, wherein the level of expression of tet operator-linked
gene can be downmodulated by administering tetracycline or a tetracycline
analogue to the mouse.
|
Claim 13
A transgenic mouse having a transgene integrated into the genome of the
mouse, wherein:
the transgene comprises a polynucleotide sequence encoding a
tetracycline-controllable transactivator fusion protein (tTA) and a
tTA-responsive promoter,
said fusion protein comprises a Tet repressor operably linked to a
polypeptide which directly or indirectly activates transcription of a gene of
interest in eucaryotic cells,
the transgene is integrated by homologous recombination at a
predetermined location within a said gene of interest within cells of the mouse
such that expression of the fission protein is controlled by 5' regulatory
elements of the gene of interest and expression of the gene of interest is
controlled by the tTA-responsive promoter,
expression of the gene of interest confers a detectable and
functional phenotype on the mouse,
said transgene is expressed in cells of the mouse at a level sufficient
to produce amounts of said fusion protein that are sufficient to activate
transcription of the gene of interest linked to the tTA-responsive promoter, and
in the absence of tetracycline or a tetracycline analogue in the mouse,
said fusion protein binds to the tTA-responsive promoter and activates
transcription of the gene of interest such that the gene of interest is
expressed at a level sufficient to confer the detectable and functional
phenotype on the mouse, wherein the level of expression of the gene of interest
can be downmodulated by administering tetracycline or a tetracycline analogue to
the mouse.
|
Claim 20
A transgenic mouse having a transgene integrated into the genome of the
mouse, wherein;
the transgene comprises a transcriptional regulatory
element functional in cells of the mouse operatively linked to a polynucleotide
sequence encoding a tetracycline-controllable transactivator fusion protein
(tTA),
said fruition protein comprises a Tet repressor operably linked to a
polypeptide which directly or indirectly activates transcription of a tet
operator-linked gene in eucaryotic cells, and
said fusion protein is expressed in cells of the mouse.
|
This patent is a Continuation in part of
US
5650298.
|
|
|
US
5866755
- Earliest priority - 14 June 1993
- Filed - 7 June 1995
- Granted - 2 February 1999
- Expected expiry - 2 February 2016
|
Title - Animals transgenic for a tetracycline-regulated
transcriptional inhibitor
Claim 1
A transgenic mouse having a transgene integrated into the genome of the mouse
and also having a tet operator-linked gene in the genome of the mouse, wherein:
the transgene comprises a transcriptional regulatory
element functional in cells of the mouse operatively linked to a polynucleotide
sequence encoding a fusion protein which inhibits transcription of said tet
operator linked gene,
said fusion protein comprises a first polypeptide
that is a Tet repressor operably linked to a heterologous second polypeptide
which inhibits transcription of said tet operator-linked gene in eucaryotic
cells,
said tet operator-linked gene confers a detectable and functional
phenotype on the mouse when expressed in cells of the mouse,
said transgene is expressed in cells of the mouse at a level sufficient
to produce amounts of said fusion protein that are sufficient to inhibit
transcription of the tet operator-linked gene; and
in the absence of tetracycline or a tetracycline analogue in the mouse,
said fusion protein binds to the tet operator-linked gene and inhibits
transcription of the tet operator linked gene, wherein the level of expression
of the tet operator-linked gene can be upregulated by administering tetracycline
or a tetracycline analogue to the mouse.
|
Claim 2
A transgenic mouse having a transgene integrated into the genome of the mouse
and also having a tet operator-linked gene in the genome of the mouse, wherein:
the transgene comprises a transcriptional regulatory
element functional in cells of the mouse operatively linked to a polynucleotide
sequence encoding a fusion protein which inhibits transcription of said tet
operator linked gene,
said fusion protein comprises a first polypeptide that is a mutated
Tet repressor that binds to tet operator sequences in the presence, but not the
absence, of tetracycline or a tetracycline analogue, operably linked to a
heterologous second polypeptide which inhibits transcription of said tet
operator-linked gene in eucaryotic cells,
said tet operator-linked gene confers a delectable and functional
phenotype on the mouse when expressed in cells of the mouse,
said transgene is expressed in cells of the mouse at a level sufficient
to produce amounts of said fusion protein that are sufficient to inhibit
transcription of the ret operator-linked gene; and
in the presence of tetracycline or a tetracycline analogue in the mouse,
said fusion protein binds to the tet operator-linked gene and inhibits
transcription of the tet operator linked gene, wherein the level of expression
of the tet operator-linked gene can be upregulated by depleting tetracycline or
a tetracycline analogue from the mouse.
|
Claim 3
A transgenic mouse having a transgene integrated into the genome of the
mouse, wherein:
the transgene comprises a transcriptional regulatory
element functional in cells of the mouse operatively linked to a polynucleotide
sequence encoding a fusion protein which inhibits transcription of a tet
operator linked gene,
the fusion protein comprising a first polypeptide
that is a Tet repressor or, a mutated Tet repressor that binds to a tet operator
sequence, operatively linked to a second polypeptide which inhibits
transcription in eukaryotic cells, and
said fusion protein is expressed in cells of the mouse.
|
This patent is a Continuation in part of
US
5789156.
|
|
|
US
5912411
- Earliest priority - 14 June 1993
- Filed - 7 June 1995
- Granted -15 June 1999
- Expected expiry - 15 June 2016
|
Title - Mice transgenic for a tetracycline-inducible
transcriptional activator
Claim 1
A transgenic mouse having a transgene integrated into the genome of the mouse
and also having a tet operator-linked gene in the genome of the mouse, wherein:
the transgene comprises a transcriptional regulatory
element functional in cells of the mouse operatively linked to a polynucleotide
sequence encoding a fusion protein which activates transcription of said tet
operator-linked gene,
the fusion protein comprises a first polypeptide
which is a mutated Tet repressor that binds to a tet operator sequence in the
presence of tetracycline or a tetracycline analogue operatively linked to a
second polypeptide which activates transcription in eukaryotic cells,
said tet operator-linked gene confers a detectable and functional
phenotype on the mouse when expressed in cells of the mouse,
said transgene is expressed in cells of the mouse at a level sufficient
to produce amounts of said fusion protein that are sufficient to activate
transcription of the tet operator-linked gene; and
in the presence of tetracycline or a tetracycline analogue in the mouse,
said fusion protein binds to the tet operator-linked gene and activates
transcription of the tet operator linked gene such that the tet operator-linked
gene is expressed at a level sufficient to confer the detectable and functional
phenotype on the mouse, wherein the level of expression of the tet
operator-linked gene cap be downmodulated by depleting tetracycline or a
tetracycline analogue from the mouse.
|
Claim 35
A transgenic mouse having a transgene integrated into the genome of the
mouse, wherein:
the transgene comprises a transcriptional regulatory
element functional in cells of the mouse operatively linked to a polynucleotide
sequence encoding a fusion protein which activates transcription of a tet
operator-linked gene,
the fusion protein comprising a first polypeptide
which is a mutated Tet repressor that binds to a tet operator sequence in the
presence of tetracycline or a tetracycline analogue operatively linked to a
second polypeptide which activates transcription in eukaryotic cells, and
said fusion protein is expressed, in cells of the mouse.
|
This patent is a Continuation in part of
US
5789156.
|
|
|
US
5922927
- Earliest priority - 14 June 1993
- Filed - 21 July 1997
- Granted -13 July 1999
- Expected expiry - 14 June 2013
|
Title - Methods for producing tetracycline-regulated
transgenic mice
Claim 1
A method for producing a transgenic mouse, comprising:
a) introducing into a fertilized oocyte of a mouse a DNA molecule
encoding a tetracycline-controllable transactivator (tTA), the tTA
comprising a prokaryotic Tet repressor operably linked to a
polypeptide which directly or indirectly activates transcription in eucaryotic
cells;
b) implanting the fertilized oocyte in a pseudopregnant foster mother;
and
c) allowing the fertilized oocyte to develop into a transgenic mouse to
thereby produce the transgenic mouse, wherein said tTA is expressed in cells of
the mouse at a level sufficient to transactivate a tet operator-linked gene.
|
Claim 2
A method for producing a transgenic mouse having a transgene encoding a
tetracycline-controllable transactivator (tTA) integrated at a predetermined
location within chromosomal DNA of cells of the mouse,
comprising:
a) introducing into a population of embryonic stem cells of a mouse a DNA
molecule encoding a tTA, the tTA comprising a prokaryotic Tet
repressor operably linked to a polypeptide which directly or indirectly
activates transcription in eucaryotic cells, the DNA molecule comprising a
polynucleotide sequence encoding the tTA flanked at 5' and 3' ends by additional
polynucleotide sequences of sufficient length for homologous recombination
between the DNA molecule and a second target DNA molecule at a predetermined
location within chromosomal DNA of cells of the mouse, under conditions suitable
for homologous recombination between the DNA molecule encoding the tTA and
chromosomal DNA within the cell;
b) selecting an embryonic stem cell in which DNA encoding the tTA has
integrated at a predetermined location within the chromosomal DNA of the cell;
c) implanting the embryonic stem cell into a blastocyst;
d) implanting the blastocyst into a pseudopregnant foster mother; and
e) allowing the blastocyst to develop into a transgenic mouse to thereby
produce the transgenic mouse, wherein said tTA is expressed in cells of the
mouse at a level sufficient to transactivate a tet operator-linked gene.
|
Claim 3
A method for producing a transgenic mouse having a transgene encoding a
tetracycline-controllable transactivator (tTA) and a tTA-responsive promoter
integrated at a predetermined location within a gene of interest in cells of the
mouse, comprising:
a) introducing into a population of embryonic stem cells of a mouse a DNA
molecule encoding a tTA, the DNA molecule comprising:
i) a first polynucleotide sequence comprising a 5'
flanking regulatory region of the gene of interest, operably linked to:
ii) a second polynucleotide sequence encoding a tTA, the tTA
comprising a prokaryotic Tet repressor operably linked to a polypeptide which
directly or indirectly activates transcription in eucaryotic cells; and
iii) a third polynucleotide sequence comprising a
tTA-responsive promoter, operably linked to:
iv) a fourth polynucleotide sequence comprising at
least a portion of a coding region of the gene of interest;
wherein the first and fourth polynucleotide sequences are of
sufficient length for homologous recombination between the DNA molecule and the
gene of interest such that expression of the tTA is controlled by 5' regulatory
elements of the gene of interest and expression of the gene of interest is
controlled by the tTA-responsive promoter, under conditions suitable for
homologous recombination between the DNA molecule encoding the tTA and the gene
of interest within the cell;
b) selecting an embryonic stem cell in which DNA encoding the tTA has
integrated at a predetermined location within the gene of interest in the cell;
c) implanting the embryonic stem cell into a blastocyst;
d) implanting the blastocyst into a pseudopregnant foster mother; and
e) allowing the blastocyst to develop into a transgenic mouse to thereby
produce the transgenic mouse,
wherein said gene of interest confers a detectable and functional
phenotype on the mouse when expressed in cells of the transgenic mouse,
said tTA is expressed in cells of the transgenic mouse at a level
sufficient to activate transcription of the gene of interest; and
in the absence of tetracycline or a tetracycline analogue in the mouse,
said tTA binds to the tTA responsive promoter operably linked to the gene of
interest and activates transcription of the gene of interest such that the gene
of interest is expressed at a level sufficient to confer the detectable and
functional phenotype on the mouse, wherein the level of expression of the gene
of interest can be downmodulated by administering tetracycline or a tetracycline
analogue to the mouse.
|
This patent is a Division of
US
5650298.
|
|
|
US
6136954
- Earliest priority - 14 June 1993
- Filed - 28 september 1998
- Granted - 24 October 2000
- Expected expiry - 14 June 2013
|
Title - Tetracycline-inducible transcriptional activator
fusion proteins
Claim 1
A fusion protein which activates transcription comprising a first polypeptide
which binds to a test operator sequence in the presence of tetracycline or a
tetracycline analogue operatively linked to a second polypeptide which activates
transcription in eukaryotic cells.
|
This patent is a Continuation of
US
5814618.
|
|
|
US
6242667
- Earliest priority - 14 June 1993
- Filed - 28 september 1998
- Granted - 5 June 2001
- Expected expiry - 14 June 2013
|
Title - Transgenic organisms having tetracycline-regulated
transcriptional regulatory systems
|
Claim 1
A transgenic plant having a transgene integrated into the genome of the plant
and also having a tet operator-linked gene in the genome of the plant, wherein:
the transgene comprises a transcriptional regulatory
element functional in cells of the plant operatively linked to a polynucleotide
sequence encoding a fusion protein which activates transcription of said tet
operator linked gene,
the fusion protein comprises a first polypeptide which is a
mutated Tet repressor that binds to a tet operator sequence in the presence of
tetracycline or a tetracycline analogue operatively linked to a second
polypeptide which activates transcription in eukaryotic cells,
said tet operator-linked gene confers a detectable and functional
phenotype on the plant when expressed in cells of the plant,
said transgene is expressed in cells of the plant at a level sufficient
to produce amounts of said fusion protein that are sufficient to activate
transcription of the tet operator-linked gene; and
in the presence of tetracycline or a tetracycline analogue in the plant,
said fusion protein binds to the tet operator-linked gene and activates
transcription of the tet operator linked gene such that the tet operator-linked
gene is expressed at a level sufficient to confer the detectable and functional
phenotype on the plant, wherein the level of expression of the tet
operator-linked gene can be downmodulated by depleting tetracycline or a
tetracycline analogue from the plant.
|
|
Claim 2
A transgenic plant having a transgene integrated into the genome of the
plant, wherein:
the transgene comprises a transcriptional regulatory
element functional in cells of the plant operatively linked to a polynucleotide
sequence encoding a fusion protein which activates transcription of a tet
operator linked gene,
the fusion protein comprising a first polypeptide which
is a mutated Tet repressor that binds to a tet operator sequence in the presence
of tetracycline or a tetracycline analogue operatively linked to a second
polypeptide which activates transcription in eukaryotic cells, and
said fusion protein is expressed in cells of the plant.
|
|
Claim 3
A transgenic plant having a transgene integrated into the genome of the plant
and also having a tet operator-linked gene in the genome of the plant, wherein:
the transgene comprises a transcriptional regulatory element functional in
cells of the plant operatively linked to a polynucleotide sequence encoding a
fusion protein which inhibits transcription of said tet operator linked gene,
the fusion protein comprises a first polypeptide which is a mutated Tet
repressor that binds to a tet operator sequence in the presence of tetracycline
or a tetracycline analogue operatively linked to a second polypeptide which
inhibits transcription in eukaryotic cells,
said tet operator-linked gene confers a detectable and functional
phenotype on the plant when expressed in cells of the plant,
said transgene is expressed in cells of the plant at a level sufficient
to produce amounts of said fusion protein that are sufficient to inhibit
transcription of the tet operator-linked gene; and
in the presence of tetracycline or a tetracycline analogue in the plant,
said fusion protein binds to the tet operator-linked gene and inhibits
transcription of the tet operator linked gene, wherein the level of expression
of the tet operator-linked gene can be upregulated by depleting tetracycline or
a tetracycline analogue from the plant.
|
|
Claim 4
A transgenic plant having a transgene integrated into the genome of the
plant, wherein:
the transgene comprises a transcriptional regulatory
element functional in cells of the plant operatively linked to a polynucleotide
sequence encoding a fusion protein which inhibits transcription of a tet
operator linked gene,
the fusion protein comprising a first polypeptide which
is a mutated Tet repressor that binds to a tet operator sequence in the presence
of tetracycline or a tetracycline analogue operatively linked to a second
polypeptide which inhibits transcription in eukaryotic cells, and
said fusion protein is expressed in cells of the plant.
|
|
Claim 5
A transgenic plant having a transgene integrated into the genome of the plant
and also having a tet operator-linked gene in the genome of the plant, wherein:
the transgene comprises a transcriptional regulatory
element functional in cells of the plant operatively linked to a polynucleotide
sequence encoding a fusion protein which inhibits transcription of said tet
operator linked gene,
said fusion protein comprises a first polypeptide that
is a Tet repressor, operably linked to a heterologous second polypeptide which
inhibits transcription of said tet operator-linked gene in eukaryotic cells,
said tet operator-linked gene confers a detectable and functional
phenotype on the plant when expressed in cells of the plant,
said transgene is expressed in cells of the plant at a level sufficient to
produce amounts of said fusion protein that are sufficient to inhibit
transcription of the tet operator-linked gene; and
in the absence of tetracycline or a tetracycline analogue in the plant,
said fusion protein binds to the tet operator-linked gene and inhibits
transcription of the tet operator linked gene, wherein the level of expression
of the tet operator-linked gene can be upregulated by administering tetracycline
or a tetracycline analogue to the plant.
|
|
Claim 6
A transgenic plant having a transgene integrated into the genome of the
plant, wherein:
the transgene comprises a transcriptional regulatory
element functional in cells of the plant operatively linked to a polynucleotide
sequence encoding a fusion protein which inhibits transcription of a tet
operator linked gene,
the fusion protein comprising a first polypeptide which
is a Tet repressor, operatively linked to a second polypeptide which inhibits
transcription in eukaryotic cells, and said fusion protein is expressed in cells
of the plant.
|
This patent is a Continuation of
US
5912411.
|
|
|
US
6252136
- Earliest priority - 14 June 1993
- Filed - 29 september 1998
- Granted - 26 June 2001
- Expected expiry - 14 June 2013
|
Title - Transgenic organisms having tetracycline-regulated
transcriptional regulatory systems
Claim 1
A transgenic plant having a transgene integrated into the genome of the plant
and also having a tet operator-linked gene in the genome of the plant,
wherein:
the transgene comprises a transcriptional regulatory
element functional in cells of the plant operatively linked to a polynucleotide
sequence encoding a fusion protein which activates transcription of said tet
operator linked gene,
the fusion protein comprises a first polypeptide which
is a Tet repressor operatively linked to a second polypeptide which directly or
indirectly activates transcription in plant cells,
said tet operator-linked gene confers a detectable and functional
phenotype on the plant when expressed in cells of the plant,
said transgene is expressed in cells of the plant at a level sufficient to
produce amounts of said fusion protein that are sufficient to activate
transcription of the tet operator-linked gene; and
in the absence of tetracycline or a tetracycline analogue in the plant,
said fusion protein binds to the tet operator-linked gene and activates
transcription of the tet operator linked gene such that the tet operator-linked
gene is expressed at a level sufficient to confer the detectable and functional
phenotype on the plant, wherein the level of expression of the tet
operator-linked gene can be downmodulated by administering tetracycline or a
tetracycline analogue to the plant.
|
Claim 7
A transgenic plant having a transgene integrated into the genome of the
plant, wherein:
the transgene comprises a transcriptional regulatory
element functional in cells of the plant operatively linked to a polynucleotide
sequence encoding a fusion protein which activates transcription of a tet
operator linked gene,
the fusion protein comprising a first polypeptide which
is a Tet repressor, operatively linked to a second polypeptide which directly or
indirectly activates transcription in plant cells, and
said fusion protein is expressed in cells of the plant.
|
This patent is a Continuation of
US
5859310.
|
|
|
US
6271348
- Earliest priority - 14 June 1993
- Filed - 24 January 2000
- Granted - 7 August 2001
- Expected expiry - 14 June 2013
|
Title - Tetracycline-inducible transcriptional inhibitor fusion proteins
|
Claim 1
A fusion protein which inhibits transcription in eukaryotic cells, the fusion
protein comprising a first polypeptide which binds to tet operator sequences,
operatively linked to a heterologous second polypeptide which inhibits
transcription in eukaryotic cells.
|
This patent is a Division of
US
6136954.
|
|
|
AU
746850 B2
- Earliest priority - 29 June 1995
- Filed - 18 August 1999
- Granted - 2 May 2002
- Expected expiry - 18 August 2019
|
Title - Tetracycline-regulated transcriptional modulaters
Claim 1
An isolated nucleic acid encoding a fusion protein which activates
transcription, the fusion protein comprising a first
polypeptide which binds to a tet operator sequence in the presence, but not the
absence, of tetracycline or a tetracycline analogue operatively linked to a
second polypeptide which activates transcription in eukaryotic cells.
|
Claim 12
A fusion protein which activates transcription comprising a
first polypeptide which binds to a tet operator sequence in the presence, but
not the absence, of tetracycline or a tetracycline or a tetracycline analogue
operatively linked to a second polypeptide which activates transcription in
eukaryotic cells.
|
Claim 35
An isolated nucleic acid encoding a fusion protein which inhibits
transcription in eukaryotic cells, the fusion protein
comprising a first polypeptide which binds to a tet operator
sequence operatively linked to a heterologous second polypeptide which inhibits
transcription in eukaryotic cells.
|
Claim 51
A fusion protein which inhibits transcription in eukaryotic cells,
comprising a first polypeptide which binds to a let operator
sequence operatively linked to a heterologous second polypeptide which inhibits
transcription in eukaryotic cells.
|
Claim 73
A host cell comprising:
a first nucleic acid encoding a first fusion protein which activates
transcription, the first fusion protein comprising a first
polypeptide which binds to a tet operator sequence operatively linked to a
second polypeptide which activates transcription in eukaryotic cells;
a second nucleic acid encoding a second fusion protein which inhibits
transcription, the second fusion protein comprising a third
polypeptide which binds to a tet operator sequence operatively linked to a
fourth polypeptide which inhibits transcription in eukaryotic cells; and
a third nucleic acid molecule comprising a nucleotide
sequence to be transcribed operatively linked to at least one tet operator
sequence.
|
Claim 76
A non-human transgenic organism comprising:
a first transgene encoding a first fusion protein which activates
transcription, the first fusion protein comprising a first
polypeptide which binds to a tet operator sequence operatively linked to a
second polypeptide which activates transcription in eukaryotic cells;
a second transgene encoding a second fusion protein which inhibits
transcription, the second fusion protein comnprising a third
polypeptide which binds to a tet operator sequence operatively linked to a
fourth polypeptide which inhibits transcription in eukaryotic cells; and
a third transgene comprising a nucleotide sequence to be
transcribed operatively linked to at least one tet operator sequence.
|
Claim 79
A recombinant vector for coordinate, bidirectional transcription of a first
and a second nucleotide sequence to be transcribed, the vector
comprising a nucleotide sequence comprising
in a 5' to 3' direction:
a first cloning site for introduction of a first nucleotide sequence to be
transcribed, which is operatively linked to at least one tet operator sequence,
which is operatively linked to a second cloning site for introduction of a
second nucleotide sequence to be transcribed, wherein transcription of the first
and second nucleotide sequence introduced into the vector proceeds in opposite
directions relative to the at least one tet operator sequence.
|
Claim 83
A composition of matter comprising at least one recombinant
vector for independent regulation of transcription of a first and a second
nucleotide sequence to be transcribed, the at least one vector
comprising a nucleotide sequence comprising:
a first cloning site for introduction of a first nucleotide sequence to be
transcribed, operatively linked to at least one tet operator sequence of a first
class type; and
a second cloning site for introduction of a second nucleotide sequence to be
transcribed, operatively linked to at least one tet operator sequence of a
second class type.
|
Claim 87
A kit comprising a carrier means having in close confinement
therein at least two container means comprising:
a first container means containing a first nucleic acid encoding a fusion
protein which activates transcription, the fusion protein
comprising a first polypeptide which binds to a let operator
sequence in the presence of tetracycline or a tetracycline analogue operatively
linked to a second polypeptide which activates transcription in eukaryotic
cells; and
a second container means containing a second nucleic acid
comprising a cloning site for introduction of a nucleotide
sequence to be transcribed operatively linked to at least one tet operator
sequence.
|
|
BASF AG
and
Abbott
GmbH & Co. KG ( a subsidiary of Abbott Labs)
|
|
Remarks
|
Related patents were granted in Canada (CA 2193122) and
Europe (
EP
804565). Related applications are pending in Europe (EP
1092771), China (CN 1167504 A and CN
1523112), Japan (JP 9500526 T2 & JP 11506901 T2)
and Norway (NO 9605623 A).
|
|
Note: patent infomation was last update on 10 May 2006. Search terms: "BASF"
in applicant and "tetracycline" in abstract. Patent database: PatentLens in
combination with INPADOC.
The protected inventions
The group of patents entitled "Tight control of gene expression in eucaryotic
cells by tetracycline-responsive promoters" are all directed to sequence
encoding a tTA fusion protein comprising a prokaryotic TetR and
a transcriptional activator domain. The claimed tTA binds to
tetO sequences in the absence of tetracycline
activating the transcription of a gene linked to a tTA
responsive promoter.
The US patent
US
5464758 claims, in addition to the tTA-encoding sequence:
- a eukaryotic cell transfected with a tTA-encoding sequence and a sequence
containing a minimal promoter with tetO sequences linked to a gene of
interest; and
- kits containing
- the tTA-encoding sequence and a second polynucleotide sequence linked to a
minimal promoter having at least a tetO sequence; and
- a eukaryotic cell transfected with the polynucleotide coding for the tTA
fusion protein.
The definition of eukaryotic cells provided by the inventors
includes yeast, plant cells, insects, mammalian and human cells.
The claims don't limit the types of regulatory sequences provided with the
tTA-encoding sequence.
The patent specification only describes constructs that are constitutively
expressed and so although at least some of the claims of this patent dominate
those of the Yale patent
(US
5851796) discussed
above, this patent does not defeat the novelty of the claims of the Yale patent.
The kit claims notably have the same peculiarities of those of the Yale
patent, e.g., requiring:
- at least two container means each containing the a different one of the two
molecules (and in one of the kit claims, the tTA-encoding molecule is already in
a cell);
- the means have to be in close confinement in the kit.
This claim format may have been copied in the Yale patent from the BASF
patent which it referenced by Application No. in the specification. It's of note
that the same law firm listed on the front page of the BASF patent is also
listed on the front page of the Yale patent.
BASF's United States patent
US
5650298 claims in addition:
- an isolated DNA sequence coding for a tTA that is recombined
with a target DNA molecule. The tTA-coding gene is flanked at each end
by DNA sequences that allow homologous recombination with the target sequence.
- In additional claims, these sequences are:
- a 5' flanking regulatory region of a gene of interest, and
- a portion of a gene of interest linked to a tTA-responsive promoter.
In this system, the expression of the tTA is controlled by the promoter of
the gene of interest and the target gene is controlled by the tTA-responsive
promoter.
Host cells (including mouse stem cells, and human cells of unrestricted type)
containing the mentioned sequences, methods for producing the host cells and
methods for producing gene products using such sequences are also claimed.
The Canadian patent CA 2165162 claims in addition to the
polynucleotide molecules of the United States patent
US
5650298:
- the use of tetracycline or a tetracycline analogue for the inhibition of the
transcription of a gene of interest linked to a tTA-responsive promoter in a
transgenic animal; and
- a method for producing a transgenic animal (non-human) by
introducing a tTA coding gene into a fertilized oocyte and implanting the oocyte
in a foster mother allowing the oocyte to develop into a transgenic animal.
The Australian patent
AU
684524 B2 claims the same type of tTA fusion protein that
recombines in a particular location in the host cell as the Canadian patent and
the United States patent
US
5650298. In addition, the Australian patent claims a
transgenic animal (non-human) having the tTA fusion protein
integrated at a predetermined location in a chromosome of the animal cells.
The United States patent
US
5589362 is directed to a sequence comprising a mutated
version of a TetR protein having at least one amino acid
mutation, which has the ability to bind a certain class (B) of
tetO sequences having a nucleotide substitution at position +4
or +6. The mutated TetR is linked to a polypeptide that regulates transcription
in eukaryotic cells. A method for regulating transcription of a class B
tetO-linked gene in a cell by introducing into the cell the fusion protein
described above.
The fusion proteins, having a first and second polypeptide, claimed in the
United States patents
US
5654168 and
US
5789156 are the opposite from each other. In the first
patent, the fusion protein works as a transcriptional
activator, whereas in the second, the fusion protein works as a
transcriptional inhibitor. The differences between the fusion
proteins are as follows:
|
Patent No.
|
First polypeptide
|
Second polypeptide
|
Tetracycline
|
Transcription
|
|
US
5654168
|
mutated TetR
|
Activator
|
presence
|
activated
|
|
US
5789156
|
TetR
|
inhibitor or silencer
|
presence
|
inhibited
|
The United States patent
US
5654168 also claims a kit carrying in separate containers:
- a tTA-encoding gene as described above; and
- a nucleotide sequence with a cloning site linked to a
tetO sequence of first class type.
The United States patent
US
5789156 also claims a kit carrying in separate containers:
- a fusion protein as described above or a eukaryotic cell
line into which the fusion protein has been introduced; and
- a nucleotide sequence with a cloning site linked to at
least a tetO sequence.
The independent claims of the patents entitled "Methods for regulating gene
expression" are methods for regulating the expression of a tetO-linked
gene in a cell. The expression is regulated by the concentration of tetracycline
administered to the cell. The differences between the
independent claims of the three United States patents are:
-
US
5814618
- in one method the fusion protein encoded by the gene introduced in the cell
contains a TetR and a second polypeptide that inhibits
transcription in the cell; and
- in a second method two different nucleotide molecules are introduced in a
cell:
- a fusion protein-encoding gene as mentioned above and
- a nucleotide sequence linked to at least a tetO sequence.
-
US
5888981
the fusion protein used in the methods claimed is a tetracycline-controllable
transactivator (tTA), which activates the
transcription in eukaryotic cells.
-
US
6004941
the TetR of the fusion protein binds to the tetO
sequences in the presence of tetracycline,
activating transcription of the gene linked to the
tetO sequences.
In addition, there are two product claims directed to recombinant
vectors containing:
- in one case, two different nucleotide sequences comprising each a cloning
site linked to the same tetO sequence for
bidirectional regulation of the transcription; and
- in another case, each of the two cloning sites is linked to different class
of tetO sequence for independent regulation of the
transcription.
The United States patents
US
5859310,
US
5866755,
US
5912411 and
US
5922927 are all directed to transgenic mice
containing a tetracycline-responsive transcriptional regulator. The transgenic
mice of the inventions have integrated in their genome:
- a transgene encoding a fusion protein, and
- a gene of interest linked to a tetO sequence.
The transcription of the gene of interest is either activated or inhibited by
the binding of the fusion protein to the tetO sequences. The fusion
protein formed by two different polynucleotides differs between the patents as
follows:
|
First polypeptide
|
Second polypeptide
|
Tetracycline
|
Transcription
|
Special feature
|
|
US
5859310
|
|
Tet repressor (TetR)
|
activator
|
absence
|
activated
|
Specific homologous recombination of tTA (fusion protein)-encoding gene and
gene of interest
|
|
US
5866755
|
|
Tet R
|
inhibitor
|
absence
|
inhibited
|
-
|
|
mutated TetR
|
inhibitor
|
presence
|
inhibited
|
-
|
|
US
5912411
|
|
mutated TetR
|
activator
|
presence
|
activated
|
-
|
|
US
5922927
|
|
prokaryotic TetR
|
activator
|
absence
|
activated
|
Specific homologous recombination of tTA (fusion protein)-encoding gene and
gene of interest
|
The United States patent
US
5922927 claims methods for producing a transgenic
mouse with a transgene coding for a fusion protein as described in the table.
Some of the methods involve the introduction of a tTA-encoding sequence in a
fertilized oocyte and its implantation in a foster mother, allowing the
development of a transgenic mouse. In other methods, the tTA-encoding gene is
introduced in embryonic stem cells of a mouse. For homologous recombination
between the tTA-encoding transgene and the gene of interest in the cells of a
mouse, the DNA molecule carrying the transgene comprises:
- a 5' regulatory region of the gene of interest linked to,
- a tTA-encoding sequence, and
- a tTA-responsive promoter linked to
- at least a portion of the gene of interest of sufficient length to mediate
homologous recombination.
The fusion protein of the United States patent
US
6136954 activates transcription in the
presence of tetracycline. A portion of the fusion protein binds
to the tetO. It's not limited to the tetR and the patent describes that mutated
tetR proteins are part of the invention.The second polypeptide of the fusion
protein is a transcriptional activator. In the broadest claims of the patent,
the activator is also not limited as to structure or amino acid sequence so long
as it functions as an activator (though the level of activation required is not
defined).
Conversely, the second polypeptide of the fusion protein claimed in the
United States patent
US
6271348 is a transcriptional inhibitor and therefore, the protein
as a whole inhibits transcription in eukaryotic cells. The
inhibitor portion also has no structural limitations in the broadest
claims. The term "inhibition" is defined in the patent as "a diminution in the
level or amount of transcription of a target gene compared to the level or
amount of transcription prior to regulation by the transcriptional inhibitor
protein." The patent describes that transcriptional inhibition may be partial
or complete.
The United States patents entitled "Transgenic organisms having tet
racycline-regulated transcriptional regulatory systems" are directed to
transgenic plants containing:
- a tetO-linked gene of interest and
- a transgene comprising a fusion protein having a first and a second
polypeptide that either activates or inhibits
the transcription of the gene linked to the tetO sequence as follows:
|
First polypeptide
|
Second polypeptide
|
Tetracycline
|
Transcription
|
|
US
6242667
|
|
mutated TetR
|
activator
|
present
|
activated
|
|
mutated TetR
|
inhibitor
|
present
|
inhibited
|
|
US
6252136
|
|
TetR
|
activator
|
absence
|
activated
|
The claims of the granted Australian patent
AU
746850 B2 are generally drawn to:
- An isolated DNA sequence coding for a fusion protein comprising:
- a first polypeptide that binds to tetO in the presence of
tetracycline, and operatively linked to
- a transcriptional activator for eukaryotic cells.
- The above fusion protein which activates transcription.
- An isolated nucleic acid sequence coding for a fusion protein where the
second polypeptide inactivates transcription.
- The above fusion protein which inhibits transcription.
- A host cell and an organism (except humans) comprising both types of fusion
proteins (activating and inhibiting transcription proteins) and a sequence of
interest to be transcribed linked to a tetO sequence.
- A recombinant vector for bidirectional transcription of
genes of interest. There are cloning sites at each end of a tetO
sequence for the introduction of the sequences to be transcribed.
- A recombinant vector for independent regulation of transcription of two
genes of interest where the tetO sequences linked to the genes are of a
different class.
- A kit comprising in separate containers
a fusion protein that activates transcription in the presence
of tetracycline and
a DNA sequence comprising a cloning site for inserting the gene
of interest linked to a tetO sequence.
The first polypeptide of the fusion proteins is not
specified in the independent claims.
The only independent claim as filed of the EP application
1092771 recites a DNA sequence encoding a fusion protein that
inhibits transcription in eukaryotic cells. The first
polypeptide of the fusion protein is defined as a TetR.
Steroid-regulated promoters
Scientific aspects
Among the chemically-inducible gene expression systems for plants and animals
are the ones based on steroid hormone receptors. The mammalian glucocorticoid
receptor (GR) is a member of the family of animal steroid
hormone receptors. GR also acts as a transcription factor by activating gene
transcription from promoters containing glucocorticoid response elements
(GRE). In the cellular environment, GR exists in a complex in
the cytoplasm with the 90-kilodaltons (kDa) heat shock protein (HSP90), which
dissociates once GR binds to its ligand (hormone).
A system comprised of the GR and GREs resulted in transient expression in
tobacco cells in the presence of dexamethasone, a strong synthetic
glucocorticoid. In stably transformed Arabidopsis plants, however, the
system did not induce expression.
The hormone binding domain (HBD) of GR and other steroid receptors can also
be used to regulate heterologous proteins in cis, that is, operatively
linked to protein-encoding sequences upon which it acts. Thus, the HBD of GR,
estrogen receptor (ER) and an insect ecdysone
receptor have shown relatively tight control and high inducibility.
The HBD of the rat GR has been linked to the DNA-binding
domain of the yeast GAL4 transcription factor and to the acidic
transactivating domain of the herpes viral protein VP16. This
chimeric transcriptional factor, named GVG, has driven the
expression of a number of genes in several plants in the presence of the ligand
dexamethasone. In some cases however, it appears to be toxic, which is caused by
a high concentration of and extended exposure to dexamethasone and induction of
untargeted genes (e.g. defense-related genes).
An ER-based inducible system for use in transgenic
plants includes a chimeric transactivator called
XVE, assembled by fusion of the regulatory region of the human
ER, the DNA-binding domain of the bacterial repressor LexA and
the acidic transactivating domain of VP16. In transgenic
tobacco and Arabidopsis, XVE has induced the
expression of genes in the presence of estradiol to levels 8-fold higher than
observed when expression is driven by a constitutive promoter, such as 35S CaMV.
The XVE system does not exhibit the toxic effects found with the GVG system, but
in some legumes (e.g., soybean) it appears to be deregulated, presumably due to
the presence of phyto-estrogens.
Since none of the systems described are induced by compounds suitable for
agricultural use, a transcription system inducible by non-steroidal
agrochemicals was developed based on the insect ecdysone
receptor. Ecdysones are insect steroidal hormones that
trigger the expression of critical genes during larval development and have been
proposed as safer alternatives to pesticides. An advantage of the ecdysone
receptor is that can also bind non-steroidal ecdysone agonists (agonists are
molecules that improve the activity of a different molecule). One hybrid
transactivator system contains the DNA-binding domain and the receptor
activation of GR and the hormone-regulatory domain of the
Heliothis virescens ER. In stable transgenic tobacco plants,
the system induced the expression of a reporter gene over 400-fold greater than
the activity of the 35S CaMV promoter. The system is highly responsive to RH5992
(tebufenozide), a non-steroidal ecdysone agonist that lacks phytotoxicity and is
used as a lepidopteran control agent on vine and horticulture plants. A drawback
of the system is a relatively high background expression.
Promoters based on glucocorticoid receptor (GR)
One of the best characterized steroid-responsive promoter systems is the GR.
It has been used widely in transgenic animal cells, in particular mammalian
cells. Granted patents claiming the basic principle of the
glucocorticoid-responsive transcriptional factors were not found. Neither were
patents or patent applications claiming a GR-based system for plants.
The system for expression in animal cells is part of an
invention disclosed in a U.S. patent granted to McGill
University.
The invention comprises an animal expression vector containing a synthetic
promoter:
- having at least 5 glucocorticoid response elements (GREs)
- GREs are placed upstream of a viral or mammalian TATA box.
- The vector also contains a transcriptional initiator site 20 to 50 bases
from the TATA box
- a restriction endonuclease site for the insertion of a gene to be expressed
from the promoter and
- lacks elements which bind to nuclear factor 1.
The promoter itself, which is also claimed, is responsive to several
steroid receptors including glucocorticoid, progesterone,
androgen and mineralocorticoid receptor. There is no limit on the types of cells
into which the promoter is transfected.
Specific Patent Information
|
Patent Number
|
Title, Independent Claims and Summary of Claims
|
Assignee
|
|
US
5512483
- Earliest priority - 21 May 1993
- Filed - 21 May 1993
- Granted - 30 April 1996
- Expected expiry - 20 May 2013
|
Title - Expression responsive to steroid hormones
Claim 1
An expression vector adapted for replication in an animal cell comprising a
glucocorticoid responsive promoter, said promoter comprising a plurality of at
least 5 glucocorticoid response elements (GREs), a viral or mammalian TATA box,
and a viral or mammalian initiator element with a transcriptional initiator site
located from 20 to 50 bases from said TATA box, said promoter lacking upstream
elements which bind nuclear factor I, and said vector further comprising a
restriction endonuclease site downstream from said promoter for insertion of DNA
to be expressed from said promoter; wherein said DNA is expressed from said
vector in an animal cell.
|
Claim 10
A promoter consisting of a plurality of at least five glucocorticoid response
elements (GREs), a TATA box, and an initiator site containing a transcriptional
initiator site located from 20 to 50 bases from said TATA box, said promoter
lacking upstream elements which bind nuclear factor I, wherein said promoter is
responsive to ligand-bound glucocorticoid, progesterone, androgen or
mineralocorticoid receptor when transiently transfected into cells, when stably
integrated within a genome, or when stably propagated in an episomal vector.
|
|
McGill University
|
|
Remarks
|
The related PCT application
WO
94/28150 and the Australian application AU 67918/94 were withdrawn in 1996.
|
Note: Patent information on this page was last updated on 21 April 2006.
Promoters based on estrogen receptor (ER)
The Rockefeller University has three granted patents in the
United States and one granted patent in Australia directed to methods for
transforming plant cells with a vector comprising a regulatory region from an
ER. It also filed two European patent applications and one
Canadian patent application.
Specific Patent Information
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Patent Number
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Title, Independent Claims and Summary of Claims
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Assignee
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US
6063985
- Earliest priority - 28 January 1998
- Filed - 28 January 1998
- Granted - 16 May 2000
- Expected expiry - 28 January 2018
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Title - Chemical inducible promoter used to obtain
transgenic plants with a silent marker
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Claim 1
A method for selecting transgenic plants comprising a silent selectable marker
wherein said method comprises the steps of: a) transforming a plant cell with a
vector wherein said vector comprises a gene selected from the group consisting
of an ipt gene, a CKI1 gene or a gene from the knotted family, wherein said gene
is under the control of an inducible promoter; b) growing said plant cells in
the absence of an exogenous plant hormone but in the presence of an exogenous
inducer of said inducible promoter; and c) excising shoots which develop,
wherein said shoots can grow into transgenic plants when grown in the absence of
said inducer.
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Independent claim 1 is drawn to:
A method for selecting transgenic plants comprising a silent selectable
marker by transforming a plant cell with a vector comprising a silent selectable
marker, selecting the plant cell in the presence of an inducer and excising
shoots for generating trasgenic plants in the absence of the inducer. The
silent selectable marker gene (an ipt gene, a CKI1 gene or a gene from the
knotted family) is under the control of an inducible promoter which can be
turned on by applying steroid (one such promoter is based on the
glucocorticoid receptor as claimed in the dependent claims).
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The Rockefeller University
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US
6784340
- Earliest priority - 28 January 1998
- Filed - 12 November 1999
- Granted - 31 August 2004
- Expected expiry - 12 November 2019
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Title - Chemical inducible promoter used to obtain
transgenic plants with a silent marker
This patent is a Continuation in part of
US
6063985.
Claim 1
A vector comprising a DNA sequence encoding a transcription factor, having
the following elements in the 5' to 3' direction, i) a promoter, ii) DNA
encoding a DNA binding domain of the bacterial repressor LexA, iii) DNA encoding
a transactivating domain of VP16, iv) DNA encoding the regulatory domain of an
estrogen receptor.
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Claim 11
An isolated nucleic acid encoding a transcription factor, comprising, in the
5' to 3' direction, i) a promoter, ii) DNA encoding a DNA binding domain of the
bacterial repressor LexA, iii) DNA encoding a transactivating domain of VP16,
iv) DNA encoding a regulatory domain of an estrogen receptor.
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US
6452068
- Earliest priority - 28 January 1998
- Filed - 12 November 1999
- Granted - 17 September 2002
- Expected expiry - 12 November 2019
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Title - Chemical inducible promoter used to obtain
transgenic plants with a silent marker
This patent is a Continuation in part of
US
6063985.
Claim 1
A method for selecting a transgenic plant encoding a silent selectable marker
wherein said method comprises the steps of: a) transforming a plant cell with a
vector wherein said vector comprises DNA encoding a regulatory region of an
estrogen receptor, a DNA-binding domain and a transactivating domain, and
further wherein said vector comprises nucleic acid encoding a silent selectable
marker which promotes shoot formation, wherein said nucleic acid is under the
control of an inducible promoter to which the DNA-binding domain binds in
response to an inducer which is estrogen or an estrogen derivative; b) growing
said plant cell in the absence of an exogenous plant hormone but in the presence
of said inducer of said inducible promoter; c) excising shoots which develop,
and d) growing said shoots to produce plants; wherein a plant produced from step
(d) is selected as being a transgenic plant.
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EP 1232273 A2
- Earliest priority - 12 November 1999
- Filed - 13 November 2000
- Granted - Pending
- Expected expiry - N/A
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Title - Chemical inducible promoters used to obtain
transgenic plants with a silent marker
Claim 1
A method for selecting transgenic plants comprising a silent selectable
marker wherein said method comprises the stepsof : a) transforming a plant cell
with a vector wherein said vector comprises DNA encoding a regulatory region of
an estrogen receptor and further wherein said vector comprises a gene which
promotes shoot formation, wherein said gene is under the control of an inducible
promoter ; b) growing said plant cells in the absence of a plant hormone but in
the presence of an inducer of said inducible promoter ; and c) excising shoots
which develop, wherein said shoots can grow into transgenic plants when grown in
the absence of said inducer.
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Claim 14
A method for inducing plant somatic embryo formation comprising the stepsof :
a) transforming a plant cell with a vector encoding a gene which promotes
somatic embryogenesis, wherein said gene is under the control of an inducible
promoter ; and b) growing said plant cells in the absence of a plant hormone but
in the presence of an inducer of said inducible promoter, wherein somatic
embryos will develop.
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Claim 26
A method for selecting transgenic plants wherein said method comprises
growing a transgenic plant, comprising an antibiotic resistance gene under the
control of a promoter comprising DNA encoding a regulatory domain of an estrogen
receptor inducible by17-P- estradiol or 4-hydroxyl tamoxifen, in the presence of
an antibiotic, wherein said antibiotic is one to which resistance is conferred
by said antibiotic resistance gene, and in the presence of17-p- estradiol or
4-hydroxyl tamoxifen.
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Claim 27
A method for selecting transgenic plants wherein said method comprises
growing a transgenic plant, comprising a herbicide resistance gene wherein said
herbicide resistance gene is under the control of a promoter comprising DNA
encoding a regulatory domain of an estrogen receptor inducible by17-P-estradiol
or 4-hydroxyl tamoxifen, in the presence of a herbicide, wherein said herbicide
is one to which resistance is conferred by said herbicide resistance gene, and
in the presence of17-P-estradiol or 4-hydroxyl tamoxifen.
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European application EP 1 232 273 A2
claims methods for selecting transgenic plants by inserting a vector having a
gene of interest under the control of an ER. When an inducer compound is added,
the transformed plants express the gene of interest. The genes under the control
of the inducible promoter are:
- genes promoting shoot formation;
- genes promoting somatic embryogenesis;
- an antibiotic resistance gene; and
- a herbicide resistance gene.
The compounds 17-beta-estradiol and 4-hydroxyl tamoxifen are cited as
inducers of the transcription system.
Although the title of the application reads chimeric promoters, these are not
subject matter of the independent claims as filed. (??- It doesn't
according to this table) However, some dependent claims mention the use
however of the DNA-binding domain of the bacterial repressor
LexA assembled with the regulatory domain of the ER.
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Remarks
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An United States application
(US
2003/150013 A1) , as a Division of
US
6452068, and a Canadian application (CA
2391003) are pending.
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EP 1242604 A2
- Earliest priority - 12 November 1999
- Filed - 13 November 2000
- Granted - Pending
- Expected expiry - N/A
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Title - Chemical inducible promoters used to obtain
transgenic plants with a silent marker and organisms and cells and methods of
using same for screening for mutations
Claim 1
A method for selecting transgenic lettuce plants comprising a silent
selectable marker wherein said method comprises the stepsof : a) transforming
lettuce root cells with a vector wherein said vector comprises a gene selected
from the group consisting of an ipt gene, aCKI1 gene, a gene from the knotted
family, and a gene the expression of which is capable of promoting shoot
regeneration, wherein said gene is under the control of an inducible promoter ;
b) growing said lettuce root cells to allow shoot development ; and c) excising
shoots which develop from plants having a shooty phenotype, wherein said shoots
can grow into normal transgenic plants when grown in the absence of said
inducer.
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Claim 11
A vector comprising a chemically inducible promoter wherein said vector
comprises
DNA encoding an estrogen receptor.
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Claim 25
A vector comprising i) a constitutive promoter, ii) DNA encoding a DNA
binding domain of bacterial repressor LexA, iii) DNA encoding a transactivating
domain of VP16, iv)DNA encoding an estrogen receptor, and v) one or more LexA
binding sites.
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Claim 26
A nucleic acid comprising a chemically inducible promoter wherein said
nucleic acid further comprises DNA encoding an estrogen receptor.
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Claim 41
A nucleic acid comprising i) a constitutive promoter, ii) DNA encoding a DNA
binding domain of bacterial repressor LexA, iii) DNA encoding a transactivating
domain of VP 16, iv) DNA encoding an estrogen receptor, and v) one or more LexA
binding sites.
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Claim 42
A transgenic lettuce plant or transgenic lettuce plant cell comprising a
vector wherein said vector comprises a chemically inducible promoter.
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Claim 43
A transgenic plant or transgenic plant cell comprising a vector wherein said
vector comprises a chemically inducible promoter which can be induced by an
estrogen.
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Claim 50
A method for making a transgenic plant display a fluorescent design, a word
or words wherein said method comprises the stepsof : a) preparing a transgenic
plant which comprises a luciferase gene under the control of a chemically
inducible promoter which is controlled by an estrogen ; and b) placing a
chemical which induces said chemically inducible promoter onto said transgenic
plant in the pattern of the design, word or words which are desired ; whereby
said plant will produce luciferase and will fluoresce in the pattern in which
the chemically inducible promoter was placed onto said transgenic plant.
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Claim 52
A transgenic lettuce plant comprising an antibiotic resistance gene wherein
said antibiotic resistance gene is under the control of an inducible promoter.
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Claim 53
A transgenic plant comprising an antibiotic resistance gene wherein said
antibiotic resistance gene is under the control of an inducible promoter,
wherein said inducible promoter comprises DNA encoding a regulatory domain of an
estrogen receptor.
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Claim 57
A transgenic plant comprising a herbicide resistance gene wherein said
herbicide resistance gene is under the control of an inducible promoter, wherein
said inducible promoter comprises DNA encoding a regulatory domain of an
estrogen receptor.
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Claim 61
An organism or a cell comprising a gene wherein a natural promoter of said
gene is lacking or inoperative and said gene is under the control of a
transgenic inducible promoter.
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Claim 74
A method to screen for mutations in a gene of an organism or cell comprising
a) preparing an organism or a cell wherein a natural promoter of said gene is
lacking or inoperative and said gene is under the control of a transgenic
inducible promoter ; and b) growing said organism or cell.
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The claims range those drawn to organisms or cells comprising a transgenic
inducible promoter to a specific chimeric inducible promoter corresponding to
the XVE promoter mentioned above in "scientific aspects" and to
lettuce plants transformed with an inducible promoter driving particular genes
promoting shoot development and antibiotic resistance.
The broadest claims originally filed recite:
- an organism or cell comprising a transgenic inducible promoter (which can be
any type of inducible promoter) which controls a gene whose natural promoter is
inoperative or lacking;
- a method for screening mutations in an organism or cell gene by putting that
gene under the control of a transgenic inducible promoter;
- a nucleic acid containing a chemically-inducible promoter having an
ER;
- a vector containing such a chemically-inducible promoter;
- transgenic plant or plant cell containing such chemically-inducible
promoter.
In other claims, the chimeric inducible promoter comprises:
- a constitutive promoter;
- a DNA-binding domain of bacterial repressor LexA;
- a transactivating domain of VP16;
- an estrogen receptor; and
- at least one LexA binding site.
In addition, the claims recite either a transgenic plant cell or specifically
a transgenic lettuce plant comprising an inducible promoter, which in some cases
explicitly comprises an ER. The same genes disclosed in EP 1282273
A2 under the control of the inducible promoter are in the claims as
filed, with the exception of genes promoting somatic embryogenesis.
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The Rockefeller University
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AU
782958 B2
- Earliest priority - 12 November 1999
- Filed - 13 November 2000
- Granted - 15 September 2005
- Expected expiry - 13 November 2020
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Title - Chemical inducible promoters used to obtain transgenic plants with a
silent marker and organisms and cells and methods of using same for screening
for mutations
Claim 1
A vector comprising a nucleic acid encoding a transcription factor, said
nucleic acid comprising in the 5' to 3' direction, i) a promoter, ii) DNA
encoding a DNA binding domain of the bacterial repressor LexA, iii) DNA encoding
a transactivating domain of VP16, and iv) DNA encoding the regulatory domain of
an estrogen receptor.
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Claim 11
A nucleic acid encoding a transcription factor, comprising, in the 5' to 3'
direction, i) a promoter, ii) DNA encoding a DNA binding domain of the bacterial
repressor LexA, iii) DNA encoding a transactivating domain of VP16, and iv) DNA
encoding a regulatory domain of an estrogen receptor.
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Note: Patent information on this page was last updated on 22 March 2006.
Promoters based on ecdysone receptor
Zeneca (now Syngenta) and Pioneer
Hi-Bred have patents and patent applications directed to:
- isolated DNA sequences of ecdysone receptors derived from diverse moth
species. Methods for inducing the expression of proteins in plants by using an
ecdysone receptor, and
- the use of other inducible receptor such as the retinoid X receptor and the
ultraspiracle receptor.
The patents granted to Syngenta protect the isolated DNA and
amino acid sequences of the ecdysone receptor from the moth species
Heliothis virescens.
Specific Patent Information
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Patent Number
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Title, Independent Claims and Summary of Claims
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Assignee
|
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US
6379945
- Earliest priority - 26 May 1995
- Filed - 24 May 1996
- Granted - 30 April 2002
- Expected expiry - 24 May 2016
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Title - Gene switch
Claim 1
An isolated or synthetic DNA sequence encoding a polypeptide selected from
the group consisting of (a) the Heliothis virescens ecdysone steroid receptor
shown in
SEQ
ID NO: 5; (b) the transactivation domain of the Heliothis virescens ecdysone
steroid receptor shown in amino acids 1-162 of
SEQ
ID NO: 5; (c) the DNA binding domain of the Heliothis virescens ecdysone
steroid receptor shown in amino acids 163-228 of
SEQ
ID NO: 5; (d) the hinge domain of the Heliothis virescens ecdysone steroid
receptor shown in amino acids 229-326 of
SEQ
ID NO: 5; e) the ligand binding domain of the Heliothis virescens ecdysone
steroid receptor shown in amino acids 327-545 of
SEQ
ID NO: 5; (f) the carboxy terminus of the Heliothis virescens ecdysone
steroid receptor shown in amino acids 546-577 of
SEQ
ID NO: 5; and (g) the hinge and ligand binding domains of the Spodoptera
exigua ecdysone steroid receptor shown in
SEQ
ID NO: 7.
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The United States patent claims an isolated or synthetic DNA
sequence encoding the mentioned ecdysone receptor and any of the following
domains of the ecdysone receptor:
- the transactivation domain;
- the DNA binding domain;
- the ligand binding domain;
- the hinge binding domain; and
- the carboxy terminus of the receptor.
Each domain has a specifically identified amino acid sequence. No particular
relationship is required between domains; e.g., the DNA sequences encoding the
individual domains can be provided in any order.
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Syngenta
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AU
711391 B2
- Earliest priority - 26 May 1995
- Filed - 20 May 1996
- Granted - 14 October 1999
- Expected expiry - 20 May 2016
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Title - A gene switch comprising an ecdysone receptor
Claim 1
DNA comprising the sequence shown in Seq. ID No. 2, or a DNA sequence which
hybridises to said DNA sequence under high stringency conditions.
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Claim 2
DNA comprising the sequence shown in Seq. ID No.3, or a DNA sequence which
hybridises to said DNA sequence under high stringency conditions.
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Claim 3
DNA comprising the sequence shown in Seq. ID No. 4, or a DNA sequence which
hybridises to said DNA sequence under high stringency conditions.
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Claim 12
A polypeptide comprising the amino acid sequence shown in Seq. ID. No. 4 or
any allelic variant or derivative thereof.
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Claim 13
A polypeptide comprising part of the amino acid sequence shown in Seq ID. No.
4 or any allelic variant or derivative thereof, which sequence provides the
Heliothis ecdysone receptor ligand binding domain.
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Claim 14
A polypeptide comprising part of the amino acid sequence shown in Seq. ID. No. 4
or any allelic variant or derivative thereof, which sequence provides the
Heliothis ecdysone receptor DNA binding domain.
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Claim 15
A polypeptide comprising part of the amino acid sequence shown in Seq. ID. No. 4
or any allelic variant or derivative thereof, which sequence provides the
Heliothis ecdysone receptor transactivation domain.
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Claim 16
A polypeptide comprising part of the amino acid sequence shown in Seq. ID. No. 4
or any allelic variant or derivative thereof, which sequence provides the
Heliothis ecdysone receptor hinge domain.
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Claim 17
A polypeptide comprising part of the amino acid sequence shown in Seq. ID. No. 4
or any allelic variant or derivative thereof, which sequence provides the
Heliothis ecdysone receptor carboxy terminal domain.
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Claim 19
DNA comprising the sequence shown in Seq. ID No.6, or a DNA sequence which
hybridises to said DNA sequence under high stringency conditions.
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The Australian patent claims both the DNA and the amino acid
sequence of Heliothis ecdysone receptor and the comprising domains of
the receptor. In this patent the species of Heliothis is not limited to
H. virescens. In addition, DNA sequences that
hybridize to the claimed sequences and any allelic
variant or derivatives of the polypeptide sequence of
the complete receptor and its domains are part of the protected invention.
The specification states that hybridization can occur at low, medium or high
stringency binding conditions. Arguably, at low stringency binding conditions,
many prior art nucleic acids will hybridize to, and therefore anticipate, this
type of claim. Similarly, a “derivative” could have a change at each nucleotide
position of the sequence and would be anticipated by many known sequences.
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EP 828829 A1
- Earliest priority - 26 May 1995
- Filed - 20 May 1996
- Granted - Pending
- Expected expiry - N/A
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Title - A gene switch comprising an ecdysone receptor
Claim 1
DNA comprising the sequence shown in Seq ID No. 2.
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Claim 2
DNA comprising the sequence shown in Seq ID No. 3.
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Claim 3
DNA comprising the sequence shown in Seq ID No. 4.
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Claim 4
DNA comprising a sequence which shows 60% or more homology with the sequence
shown in Seq ID No.1, 2 or 3.
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Claim 6
DNA which hybridises to the sequence shown in Seq ID No.2, 3 or 4, and which
codes for at leaset part of the Heliothis ecdysone receptor.
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Claim 8
DNA comprising part of the sequece shown in Seq ID No.2, and which codes for
at least part of the Heliothis ecdysone receptor ligand binding domain.
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Claim 9
DNA comprising part of the sequece shown in Seq ID No.3, and which codes for
at least part of the Heliothis ecdysone receptor ligand binding domain.
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Claim 10
DNA comprising part of the sequece shown in Seq ID No.4, and which codes for
at least part of the Heliothis ecdysone receptor ligand binding domain.
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Claim 15
DNA comprising part of the sequece shown in Seq ID No.2, and which codes for
at least part of the Heliothis ecdysone receptor DNA binding domain.
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Claim 16
DNA comprising part of the sequece shown in Seq ID No.3, and which codes for
at least part of the Heliothis ecdysone receptor DNA binding domain.
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Claim 17
DNA comprising part of the sequece shown in Seq ID No.4, and which codes for
at least part of the Heliothis ecdysone receptor DNA binding domain.
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Claim 22
DNA comprising part of the sequece shown in Seq ID No.2, and which codes for
at least part of the Heliothis ecdysone receptor transactivation
domain.
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Claim 23
DNA comprising part of the sequece shown in Seq ID No.3, and which codes for
at least part of the Heliothis ecdysone receptor transactivation
domain.
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Claim 24
DNA comprising part of the sequece shown in Seq ID No.4, and which codes for
at least part of the Heliothis ecdysone receptor transactivation
domain.
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Claim 29
DNA comprising part of the sequece shown in Seq ID No.2, and which codes for
at least part of the Heliothis ecdysone receptor hinge domain.
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Claim 30
DNA comprising part of the sequece shown in Seq ID No.3, and which codes for
at least part of the Heliothis ecdysone receptor hinge domain.
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Claim 31
DNA comprising part of the sequece shown in Seq ID No.4, and which codes for
at least part of the Heliothis ecdysone receptor hinge domain.
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Claim 36
DNA comprising part of the sequece shown in Seq ID No.2, and which codes for
at least part of the Heliothis ecdysone receptor carboxy terminal
region.
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Claim 37
DNA comprising part of the sequece shown in Seq ID No.3, and which codes for
at least part of the Heliothis ecdysone receptor carboxy terminal
region.
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Claim 38
DNA comprising part of the sequece shown in Seq ID No.4, and which codes for
at least part of the Heliothis ecdysone receptor carboxy terminal
region.
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Claim 44
A polypeptide comprising the amino acid sequence shown in Seq ID No.4 or any
allelic variant or derivative thereof.
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Claim 45
A polypeptide comprising part of the amino acid sequence shown in Seq ID No.4 or
any allelic variant or derivative thereof, which sequence provides the
Heliothis ecdysone receptor ligand binding domain.
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Claim 46
A polypeptide comprising part of the amino acid sequence shown in Seq ID No.4 or
any allelic variant or derivative thereof, which sequence provides the
Heliothis ecdysone receptor DNA binding domain.
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Claim 47
A polypeptide comprising part of the amino acid sequence shown in Seq ID No.4 or
any allelic variant or derivative thereof, which sequence provides the
Heliothis ecdysone receptor transactivation domain.
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Claim 48
A polypeptide comprising part of the amino acid sequence shown in Seq ID No.4 or
any allelic variant or derivative thereof, which sequence provides the
Heliothis ecdysone receptor hinge domain.
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Claim 49
A polypeptide comprising part of the amino acid sequence shown in Seq ID No.4 or
any allelic variant or derivative thereof, which sequence provides the
Heliothis ecdysone receptor carboxy terminal region.
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Claim 51
DNA comprising the sequence shown in Seq ID No. 6.
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Claim 52
DNA comprising a sequence which shows 60% or more homology with the sequence
shown in Seq ID No.6.
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Claim 54
DNA which hybridises to the DNA sequence shown in Seq ID No. 6 and which codes
for at least part of Spodoptera ecdysone receptor.
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Claim 56
DNA comprising part of the sequece shown in Seq ID No.6, and which codes for
at least part of the Spodoptera ecdysone receptor ligand binding
domain.
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Claim 61
DNA comprising part of the sequece shown in Seq ID No.6, and which codes for
at least part of the Spodoptera ecdysone receptor hinge domain.
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The claims of the European patent application, in addition, recite DNA
sequences encoding a Spodoptera ecdysone receptor from the moth genus
Spodoptera.
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Remarks
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Related applications also filed in Canada (CA 2219121), Bulgaria (BG 102124
A), Brazil (BR 9608897 A), Czech Republic (CZ9703722 A3), Hungary (HU 9802225
AB), Japan (JP 11506319 T2), Norway (NO 975419 A), New Zealand (NZ 308162 A),
Poland (PL 323587 A1) and Turkey (TR 9701436 T1). The application in China (CN
1191568 AA) has lapsed.
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EP 1112360 A1
- Earliest priority - 10 September 1998
- Filed -10 September 1999
- Granted - Pending
- Expected expiry - N/A
|
Title - Ecdysone receptors and methods for their use
|
Claim 1
An isolated nucleic acid sequence that encodes an insect protein from a
Pyrilidae species, wherein said nucleotide sequence is selected from the group
consisting of
a) a nucleotide sequence comprising a sequence encoding an ecdysone receptor
or Ultraspiracle;
b) a nucleotide sequence comprising the sequence set forth in SEQ ID NO:1 or
3;
c) a nucleotide sequence encoding a polypeptide comprising the amino acid
sequence set forth in SEQ ID NO: 2 or 4;
d) a nucleotide sequence that hybridizes under stringent conditions to a
sequence of a), b), or c).
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|
Claim 2
An isolated polypeptide from a Pyrilidae species, wherein said polypeptide is
selected from the group consisting of
a) a polypeptide sequence comprising an Ecdysone receptor or Ultraspiracle;
b) a polypeptide comprising an amino acid sequence set forth in SEQ ID NO:2
or 4;
c) a polypeptide encoded by a nucleotide sequence comprising the sequence
set forth in SEQ ID NO:1 or 3;
d) a polypeptide encoded by a nucleotide sequence that hybridizes under
stringent conditions to a nucleotide sequence comprising the sequence set forth
in SEQ ID NO:1 or 3.
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Claim 3
A method of selectively inducing gene expression of a protein of interest in a
plant, said method comprising:
a) stably incorporating into the genome of said plant an expression
cassette, said expression cassette comprising a promoter operably linked to a
nucleotide sequence encoding an Ecdysone receptor, wherein said nucleotide
sequence encoding the Ecdysone receptor is selected from the group consisting of
i) a nucleotide sequence from a Pyrilidae species comprising a sequence
encoding an Ecdysone receptor;
ii) a nucleotide sequence comprising the sequence set forth in SEQ ID
NO:1;
iii) a nucleotide sequence encoding a polypeptide comprising the amino
acid sequence set forth in SEQ ID NO: 2;
iv) a nucleotide sequence that hybridizes under stringent conditions to
a sequence of i), ii), or iii);
b) further stably incorporating into the genome of said plant a second
expression cassette, wherein said expression cassette comprises a
transcriptional regulatory region operably linked to a nucleotide sequence
encoding said protein of interest, and wherein said transcriptional regulatory
region is activated by the ligand-receptor complex;
c) contacting said plant with a ligand which complexes with said receptor,
wherein said receptor-ligand complex interacts with the transcription regulatory
region and induces gene expression of the protein of interest.
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|
Claim 23
An expression vector comprising a promoter operably linked to a nucleotide
sequence encoding an Ecdysone receptor wherein said nucleotide sequence encoding
the Ecdysone receptor is selected from the group consisting of:
a) a nucleotide sequence from a Pyrilidae species comprising a sequence
encoding an Ecdysone receptor;
b) a nucleotide sequence comprising the sequence set forth in SEQ ID NO:1;
c) a nucleotide sequence encoding a polypeptide comprising the amino acid
sequence set forth in SEQ ID NO: 2;
d) a nucleotide sequence that hybridizes under stringent conditions to a
sequence of a), b), or c).
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|
Claim 25
An expression vector comprising a promoter operably linked to a nucleotide
sequence encoding Ultraspiracle, and said nucleotide sequence encoding
Ultraspiracle is selected from the group consisting of
a) a nucleotide sequence comprising the sequence set forth in SEQ ID NO:3;
b) a nucleotide sequence encoding a polypeptide comprising the amino acid
sequence set forth in SEQ ID NO: 4;
c) a nucleotide sequence that hybridizes under stringent conditions to a
sequence of a) or b).
|
The claims of the European patent application recite the
isolated DNA sequences and the polypeptide sequences of an ecdysone
receptor and an ultraspiracle derived from insects of
the family Pyralidae, which is a moth family. Ecdysone response
elements found in the promoters of the target genes are activated by
heterodimers formed by an ultraspiracle and the ecdysone receptor.
The claims as filed also recite methods of inducing gene expression in a
plant where the gene of interest is linked to a transcriptional regulatory
region that responds to the ecdysone receptor-ligand complex and is activated by
the complex. Expression vectors comprising a promoter operably linked to an
ecdysone receptor encoding sequence and an ultraspiracle are also part of the
disclosed invention.
|
Pioneer Hi-Bred International Inc.
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Remarks
|
Application also filed in Australia (AU 59159/99 A1)
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Note: Patent information on this page was last updated on 7 April 2006.
Other patents related to steroid-responsive promoters
There is an increasing number of patent documents related to the
steroid/retinoid/thyroid receptor superfamily and their multiple applications,
which are beyond of the scope of this report. The disclosure of the European
application EP 1112360 A1 discusses several patent applications
and patents related to this topic. As such, it is a good reference to those who
would like to assess the patent landscape in this field in a much broader
context.
The following list of patent documents refers to chimeric inducible receptors
that combine domains from steroid-responsive promoters such as glucocorticoid,
mineralocorticoid, and estrogen, among others. Some of them also combine
receptors that can respond to both steroid and metal compounds making it a
doubly inducible promoter system. Finally, ecdysone receptors isolated from
different insects and new receptors based on the retinoic acid receptor are also
included. In no way do these documents represent the total number of patents
directed to this extensive topic, but they give an idea of the diverse
applications in this field of technology.
|
Patent/application number
|
Title, Independent Claims and Summary of
Claims
|
Assignee
|
|
WO
01/62780
- Earliest priority - 24 February 2000
- Filed - 30 August 2001
|
Title - Gene expression system based on chimeric receptors
Claim 1
A system for modulating the expression of a target gene associated with a
defined response element in a subject, said system comprising:
a first chimeric protein comprising at least one
dimerization domain of a first member of the steroid/thyroid hormone nuclear
receptor superfamily and at least one DNA binding domain, and
a second chimeric protein comprising at least one
dimerization domain of a second member of the steroid/thyroid hormone nuclear
receptor superfamily and at least one transcription modulating domain, wherein
the first and second chimeric proteins associate to form a functional entity
under substantially physiological conditions, and wherein response of said
response element to said DNA binding domain modulates expression of said target
gene.
|
|
Claim 27
A system for modulating the expression of a target gene associated with a
defined response element in a subject, said system comprising:
a first chimeric protein comprising at least one
dimerization domain of a first member of the steroid/thyroid hormone nuclear
receptor superfamily and at least one DNA binding domain, and
a second chimeric protein comprising at least one
dimerization domain of a second member of the steroid/thyroid hormone nuclear
receptor superfamily and at least one transcription modulating domain, and
wherein, at least one of the receptors is non-endogenous to said subject and the
first and second chimeric proteins associate to form a functional entity under
substantially physiological conditions in the presence or absence of a
non-endogenous ligand, and wherein response of said response element to said DNA
binding domain modulates expression of said target gene.
|
|
Claim 46
A system for modulating the expression of a target gene target gene associated
with a defined response element in a subject, said system
comprising:
a first chimeric protein consisting of a DNA binding domain
and a dimerization domain of a first member of the steroid/thyroid hormone
nuclear receptor superfamily, and
a second chimeric protein consisting of a transcription
modulating domain and a dimerization domain of a second member of the
steroid/thyroid hormone nuclear receptor superfamily, wherein at least one of
the receptors is non-endogenous and the first and second chimeric proteins
associate to form a functional entity under substantially physiological
conditions in the presence of a non-endogenous ligand, and wherein association
of said response element with said DNA binding domain modulates expression of
said target gene.
|
The claims are geneally directed to systems to modulate the expression of a
target gene by the interaction of two chimeric proteins each having a
dimerization domain member of the streoid/thyroid hormone nuclear receptor. The
dimerization domain is required for functional transactivation. The chimeric
proteins also contain a DNA-binding domain and a transcription-modulating
domain.
|
The Salk Institute for Biological Studies
|
|
Remarks
|
Related application in the United States (
US
2004/102367) is pending, and applications in Australia (AU
2001241682) and Europe (EP 1259537) were lapsed and
withdrawn, respectively.
|
|
US
6723531
- Earliest priority - 5 April 1996
- Filed - 16 March 1998
- Granted - 20 April 2004
- Expected expiry - 5 April 2016
|
Title - Methods for modulating expression of exogenous genes in mammalian
systems, and products related thereto
Claim 1
A method for modulating the expression of an exogenous gene in an isolated
cell containing:
(i) a modified ecdysone receptor which, in the presence of a ligand
therefor, and optionally in the further presence of a silent partner therefor,
binds to a response element, wherein said modified ecdysone receptor
comprises:
(a) a ligand binding domain that binds to an ecdysteroid,
(b) a DNA-binding domain obtained from a DNA-binding protein, which
binds to said response clement; and
(c) an activation domain of a transcription factor, wherein at least one
of said DNA-binding domain or said activation domain is not obtained from a
native ecdysone receptor, with the proviso that when said activation domain is
derived from a glucocorticoid receptor, said DNA-binding domain is not derived
from a glucocorticoid receptor or a E. coli LexA protein; and
(ii) a DNA construct comprising said exogenous gene under
the control of said response element, wherein said response element:
(a) is a modified response element which comprises, in any order, a
first half-site and a second half-site separated by a spacer of 0-5 nucleotides;
wherein said first half-site has the sequence: EQU --RGBNNM--, wherein each R is
independently selected from A or G; each B is independently selected from G, C,
or T; each N is independently selected from A, T, C, or G; and each M is
independently selected from A or C; with the proviso that at least 4 nucleotides
of each --RGBNNM--group of nucleotides are identical with the nucleotides at
comparable positions of the sequence --AGGTCA--; and wherein said second
half-site is obtained from a glucocorticoid receptor subfamily response element,
(b) binds to said modified ecdysone receptor, and
(c) does not bind to farnesoid X receptor (FXR); said method comprising
providing to the cell an effective amount of one or more ligands for said
modified ecdysone receptor; wherein said one or more ligands are not normally
present in the cell; and wherein said one or more ligands are not toxic to said
cell.
|
Claim 18
A method of inducing the expression of an exogenous gene in an isolated cell
containing:
(i) DNA encoding a modified ecdysone receptor under the control of an
inducible promoter, wherein said modified ecdysone receptor, in the presence of
a ligand therefor, and optionally in the further presence of a silent partner
therefor, binds to a response element, and wherein said modified ecdysone
receptor comprises:
(a) a ligand binding domain that binds to an ecdysteroid,
(b) a DNA-binding domain obtained from a DNA-binding protein, which
binds to said response element; and
(c) an activation domain of a transcription factor, wherein at least one
of said DNA-binding domain or said activation domain is not obtained from a
native ecdysone receptor, with the proviso that when said activation domain is
derived from a glucocorticoid receptor, said DNA-binding domain is not derived
from a glucocorticoid receptor or an E. coli LexA protein;
(ii) a DNA construct comprising said exogenous gene under
the control of said response element, wherein said response elements;
(a) is a modified response element which comprises, in any order, a
first half-site and a second half-site separated by a spacer of 0-5 nucleotides;
wherein said first half-site has the sequence: EQU --RGBNNM--, wherein each R is
independently selected from A or G; each B is independently selected from G, C,
or T; each N Is independently selected from A, T, C, or G; and each M is
independently selected from A or C; with the proviso that at least 4 nucleotides
of each --RGBNNM--group of nucleotides are identical with the nucleotides at
comparable positions of the sequence --AGGTCA--; and wherein said second
half-site is obtained from a glucocorticoid receptor subfamily response element,
(b) binds to said modified ecdysone receptor, and
(c) does not bind to farnesoid X receptor (FXR); and (iii) one or more
ligands for said modified ecdysone receptor; said method
comprising subjecting said cell to conditions suitable to
induce expression of said modified ecdysone receptor.
|
Claim 19
A method of inducing expression of an exogenous gene in an isolated cell
containing a DNA construct containing said
exogenous gene under the control of a response element, wherein said response
element:
(a) is a modified response element which comprises, in any order, a first
half-site and a second half-site separated by a spacer of 0-5 nucleotides;
wherein said first half-site has the sequence; EQU --RGBNNM--, wherein each R is
independently selected from A or G; each B is independently selected from G, C,
or T; each N is independently selected from A, T, C, or G; and each M is
independently selected front A or C; with thy proviso that at least 4
nucleotides of each --RGBNNM--group of nucleotides are identical with the
nucleotides at comparable position of the sequence --AGGTCA--; and wherein said
second half-site is obtained from a glucocorticoid receptor subfamily response
element,
(b) binds to said modified ecdysone receptor, and
(c) does not bind to farnesoid X receptor (FXR), said method
comprising introducing into said cell a modified ecdysone
receptor, wherein said modified ecdysone receptor comprises:
(a) a ligand binding domain that binds to an ecdysteroid,
(b) a DNA-binding domain obtained from a DNA-binding protein, which binds to
said response element; and
(c) an activation domain of a transcription factor, wherein at least one of
said DNA-binding domain or said activation domain is not obtained from a native
ecdysone receptor, with the proviso that when said activation domain is derived
from a glucocorticoid receptor, said DNA-binding domain is not derived from a
glucocorticoid receptor or an E. coli LexA protein; and one or more ligands for
said modified ecdysone receptor, wherein said receptor, in combination with a
ligand therefor, and optionally in the further presence of a silent partner
therefor, binds to said response element, activating transcription therefrom.
|
|
Claim 20
A method for the expression of a recombinant product detrimental to isolated
host cells, said method comprising: transforming suitable
isolated host cells with:
(i) DNA encoding a modified ecdysone receptor, wherein said modified
ecdysone receptor comprises:
(a) a ligand binding domain that binds to an ecdysteroid,
(b) a DNA-binding domain obtained from a DNA-binding protein; and
(c) an activation domain of a transcription factor, wherein at least one
of said DNA-binding domain or said activation domain is not obtained from a
native ecdysone receptor, with the proviso that when said activation domain is
derived from a glucocorticoid receptor, said DNA-binding domain is not derived
from a glucocorticoid receptor or an E. coli LexA protein; and
(ii) a DNA construct encoding said recombinant product under the control of
a response element, wherein said response element;
(a) is a modified response element which comprises, in any order, a
first half-site and a second half-site separated by a spacer of 0-5 nucleotides;
wherein maid first half-site has the sequence; EQU --RGBNNM--, wherein each R is
independently selected from A or G; each B is independently selected from G, C,
or G; each N is independently selected from A, T, C, or G; and each M is
independently selected from A or C; with the proviso that at least 4 nucleotides
of each --RGBNNM--group of nucleotides are identical with the nucleotides at
comparable positions of the sequence --AGGTCA--; and wherein said second
half-site is obtained from a glucocorticoid receptor subfamily response element
(b) binds to said modified ecdysone receptor, and
(c) does not bind to farnesoid X receptor (FXR); growing said host cells
in suitable media; and inducing expression of said recombinant product by
introducing into said host cells one or more ligands for said modified ecdysone
receptor, and optionally a silent partner for said modified ecdysone receptor.
|
The claims are drawn to methods for modulating the expression of an exogenous
gene in a mammalian cell by using modified ecdysone receptors that is paired
with a silent partner of the steroid/thyroid superfamily of receptors.
|
|
Remarks
|
Related patent is granted in Australia
(AU
734051 B2). Applications are also pending in Europe (EP
910652 A1) and Canada (CA 2251466)
|
|
US
5599904
- Earliest priority - 2 December 1987
- Filed - 3 March 1992
- Granted - 4 February 1997
- Expected expiry - 4 February 2014
|
Title - Chimeric steroid hormone superfamily receptor proteins
Claim 1
A chimeric receptor having at least an N-terminus domain, a
DNA-binding domain, and a ligand-binding domain; wherein each of said
N-terminus, DNA-binding and ligand-binding domains is obtained from a member of
the steroid hormone superfamily of receptors; wherein said domains of said
chimeric receptor originate from at least two different members of the steroid
hormone superfamily of receptors; and wherein at least one of the domains of
said chimeric receptor originates from a human retinoic acid receptor alpha.
|
Claim 2
A chimeric receptor having at least a DNA-binding domain and
a ligand-binding domain; wherein each of said DNA-binding and ligand-binding
domains is obtained from a member of the steroid hormone superfamily of
receptors; wherein said domains of said chimeric receptor originate from at
least two different members of the steroid hormone superfamily of receptors; and
wherein at least one of the domains of said chimeric receptor originates from a
human retinoic acid receptor alpha.
|
The claims are to chimeric receptor having component domains derived from at
least two different members of the steroid hormone superfamily of receptors,
with one of the domains derived from a human retinoic acid receptor alpha.
|
|
US
4981784
- Earliest priority - 2 December 1987
- Filed - 30 November 1988
- Granted - 1 January 1991
- Expected expiry - 30 November 2008
|
Title - Retinoic acid receptor composition and method for identifying ligands
Claim 1
A method for identifying functional ligands for receptor proteins, said
method comprising:
(a) isolating DNA sequences having a ligand-binding domain and a DNA-binding
domain;
(b) constructing a chimeric gene by substituting operative portions of the
DNA-binding domain region of the DNA sequence of step (a) with operative
portions of a DNA-binding domain region from a known ligand-responsive receptor
protein;
(c) introducing into a suitable receptor-deficient host cell:
(1) the chimeric gene from step (b), and
(2) a reporter gene functionally linked to an operative hormone response
element wherein the hormone response element is capable of being activated by
the DNA-binding domain region of the receptor protein encoded by the chimeric
gene of step (b); (d) challenging the transfected host cell from step (c)
with at least one compound to be evaluated for ligand bidning activity with the
chimeric receptor protein encoded by the chimeric gene of step (b);
(e) monitoring induction of the reporter gene;
(f) identifying as a functional ligand(s) that ligand(s) which is capable of
inducing production of the protein product of the reporter gene.
|
Claim 7
A method for identifying functional ligands for receptor proteins in a cell
wherein said cell contains,
(a) an expressible chimeric DNA sequence (c) comprised
of operative portions of a DNA-binding domain of a first
receptor sequence linked to operative portions of a ligand-binding domain of a
second receptor sequence, and
(b) a reporter nucleic acid sequence functionally linked to an operative
hormone response element wherein said chimeric DNA sequence is expressed and
wherein the DNA-binding domain of the chimeric receptor protein thus produced
can functionally bind to and activate the hormone response element that is
functionally linked to the reporter sequence, said method
comprising challenging the cell with at least one compound to
be evaluated for ligand binding activity wherein said compound to be evaluated
is not known to be a functional ligand for the chimeric protein encoded by said
chimeric DNA sequence (c).
|
The patent mainly claims for methods for identifying ligands for receptor
proteins by replacing the DNA-binding domain of a putative novel receptor with
the DNA-binding domain of a known receptor. The expression of the hybrid
receptor depends on the presence of the new ligand.
|
|
Remarks
|
Related patent was also granted in Europe
(EP
325849 B1).
|
|
US
5646013
- Earliest priority - 9 November 1988
- Filed - 28 June 1995
- Granted - 8 July 1997
- Expected expiry - 9 November 2008
|
Title - Method of producing foreign products
Claim 1
A method of producing gene products in mammalian cells
comprising:
(1) transfecting mammalian cells with both:
(a) a first expression plasmid, comprising a mouse
mammary tumor virus (MMTV) long terminal repeat (LTR) operably linked to a
glucocorticoid receptor protein gene (GRP), wherein on transfection said cells
produce glucocorticoid receptor protein as a result of expression of said first
expression plasmid; and
(b) a second expression plasmid, comprising an MMTV LTR
operably linked to a gene encoding a physiologically active substance, wherein
on transfection said cell produces said physiologically active substance as a
result of expression of said second expression plasmid; wherein on exposing the
resulting transfected cells to a glucocorticoid, transfected mammalian cells
that exhibit greater expression of said physiologically active substance in the
presence of said glucocorticoid than in the absence thereof are selected;
(2) propagating the thus-obtained transfected cells in the presence of said
glucocorticoid to induce expression of said physiologically active substance;
and
(3) obtaining said physiologically active substance.
|
Claim 2
A method of producing foreign gene products in mammalian cells bearing, on a
chromosome thereof:
(a) a first mouse mammary tumor virus (MMTV) long terminal repeat (LTR)
operably linked to a glucocorticoid receptor protein gene (GRP), wherein on
transfection said cells produce glucocorticoid receptor protein as a result of
expression of said first MMTV LTR operably linked to said GRP gene; and
(b) a second MMTV LTR operably linked to a gene encoding a physiologically
active substance, wherein on transfection said cells produce said
physiologically active substance as a result of expression of said second MMTV
LTR operably linked to said gene encoding said physiologically active substance;
wherein on exposing the resulting transfected cells to a glucocorticoid,
transfected mammalian cells that exhibit greater expression of said
physiologically active substance in the presence of said glucocorticoid than in
the absence thereof are selected, comprising:
(1) propagating said cells in the presence of said glucocorticoid to
induce expression of said physiologically active substance; and
(2) obtaining said physiologically active substance.
|
The claims of this patent are directed to methods using inducible promoter
system for mammalian cells comprising a first plasmid having a mouse mammary
tumor virus (MMTV) long terminal repeat (LTR) linked to a glucocorticoid
receptor and a second plasmid with a MMTV-LTR linked to a gene of interest. The
plasmids interact for the expression of the gene of interest in the presence of
a glucocorticoid.
|
Daiichi Seiyaku Co. Ltd
|
|
Remarks
|
Related patent was also granted in Europe
(EP
316717 B1).
|
|
US
5877018
- Earliest priority - 20 April 1994
- Filed - 16 April 1996
- Granted - 2 March 1999
- Expected expiry - 20 April 2014
|
Title - Synthetic eukaryotic promoters containing two inducible elements
Claim 1
A synthetic inducible eukaryotic promoter for the regulation of transcription
of a gene, comprising at least two different classes of inducible elements,
wherein said different classes of inducible elements are selected to provide a
synergistic level of expression of a gene product in a eukaryotic expression
system, said promoter being derived from a native promoter containing at least
one constitutive element, wherein said at least one constitutive element is
functionally disabled to decrease the level of basel gene expression by said
promoter and to increase the ratio of induced to basal gene expression by said
promoter.
|
Claim 3
A synthetic inducible eukaryotic promoter for the regulation of transcription
of a gene, comprising at least two different classes of inducible elements,
wherein said different classes of inducible elements are selected to provide a
synergistic level of expression of a gene product in a eukaryotic expression
system, said promoter being derived from a native promoter and one of said
different classes of inducible elements is a native inducible element and
another of said different classes of inducible elements is a different inducible
element provided in said native promoter, wherein said native inducible element
is a metal-responsive element (MRE) and said different inducible element is at
least one glucocorticoid-responsive element (GRE) and is provided in said native
promoter by insertion.
|
Claim 12
A synthetic inducible eukaryotic promoter for the regulation of transcription
of a gene, comprising at least two different classes of inducible elements,
wherein said different classes of inducible elements are selected to provide a
synergistic level of expression of a gene product in a eukaryotic expression
system, said promoter being derived from a native promoter and one of said
different classes of inducible elements is a native inducible element and
another of said different classes of inducible elements is a different inducible
element provided in said native promoter, wherein said native inducible element
is a metal responsive element (MRE) and said different inducible element is a
glucocorticoid-responsive element (GRE) and is provided in said native promoter
by insertion, wherein said native promoter is the hMT-IIA promoter, wherein
multiple linked GRE's are inserted into the native promoter.
|
Claim 19
A synthetic inducible eukaryotic promoter for the regulation of transcription
of a gene, comprising at least two different classes of inducible elements,
wherein said different classes of inducible elements are selected to provide a
synergistic level of expression of a gene product in a eukaryotic expression
system, said promoter being derived from a native promoter and one of said
different classes of inducible elements is a native inducible element and
another of said different classes of inducible elements is a different inducible
element provided in said native promoter, wherein said native responsive element
is a glucocorticoid-responsive element (GRE) and said different inducible
element is a metal responsive element (MRE) which is provided in said native
promoter by insertion.
|
Claim 23
A synthetic inducible eukaryotic promoter for the regulation of transcription
of a gene, comprising at least two different classes of inducible elements,
wherein said different classes of inducible elements are selected to provide a
synergistic level of expression of a gene product in a eukaryotic expression
system, said promoter being derived from a native promoter and one of said
different classes of inducible elements is a element is a native inducible
element and another of said different classes of inducible elements is a
different inducible element provided in said native promoter, wherein said
native responsive element is a glucocorticoid-responsive element (GRE) and said
different inducible element is a metal responsive element (MRE) which is
provided in said native promoter by insertion, wherein said native promoter is
the MMTV-LTR promoter, wherein at least two linked MRE's are inserted into the
native promoter.
|
- Synthetic inducible promoters containing a metal-responsive element and a
glucocorticoid responsive element for driving the expression of a gene in a
eukaryotic system.
Inducible eukaryotic promoters derived from a human metallothionen gene and
mouse mammary tumor virus. The first one contains metal-responsive elements and
the second one glucocorticoid-responsive elements.
|
Connaught Laboratories Ltd
|
|
US
5559027
- Earliest priority - 30 March 1993
- Filed - 20 October 1994
- Granted - 24 September 1996
- Expected expiry - 24 September 2013
|
Title - Synthetic eukaryotic promoters containing two inducible elements
Claim 1
A synthetic inducible eukaryotic promoter for the regulation of transcription
of a gene to produce a gene product, comprising at least two inserted linked
responsive elements from a different class of inducible promoter, wherein (a)
the inducible promoter is selected from the group consisting of the human
metallothionein gene (hMT-IIA) promoter and the mouse mammary tumor virus/long
terminal repeat (MMTV-LTR)promoter, (b) the inserted responsive elements are
selected from the group consisting of glucocorticoid-responsive elements (GREs)
and metal-responsive elements (MREs), and (c) said different classes of
inducible elements are inserted such that induction of the promoter by both
classes of inducers causes a synergistic increase in expression of the gene
product in a eukaryotic expression system compared to induction by either
inducer alone.
|
|
|
EP
633941 B1
- Earliest priority - 30 March 1992
- Filed - 30 March 1992
- Granted - 15 May 2002
- Expected expiry - 30 March 2012
|
Title - Synthetic eukaryotic promoters containing two inducible elements
Claim 1
A synthetic inducible eukaryotic promoter for the regulation of transcription
of a gene, comprising at least two different classes of inducible elements,
wherein said different classes of inducible elements are selected to provide a
synergistic level of expression of a gene product in a eukaryotic expression
system, characterised in that said classes of inducible elements are selected
from the group consisting of hormone-responsive elements (HREs),
metal-responsive elements (MREs), heat shock-responsive elements (HSREs) and
interferon-responsive elements (IREs).
|
|
|
US
5514578
- Earliest priority - 26 February 1990
- Filed - 30 September 1992
- Granted - 7 May 1996
- Expected expiry - 7 May 2013
|
Title - Polynucleotides encoding insect steroid hormone receptor polypeptides
and cells transformed with same. / Polynucleotide encoding insect ecdysone
receptor
Claim 1
An isolated polynucleotide encoding a Drosophila ecdysone receptor having the
878 amino acid predicted amino acid sequence of FIG. 4 (A-C) and which has
ecdysteroid-binding activity wherein said ecdysone receptor exhibits saturable
binding to β-ecdysone or ecdysone analogues.
|
Claim 5
A transformed insect bacterial or mammalian host cell comprising a
polynucleotide encoding a Drosophila ecdysone receptor consistinq of the 878
amino acid predicted amino acid sequence of FIG. 4 (A-C) operably linked to a
promoter, wherein said promoter is not naturally associated with an ecdysone
receptor gene in the germline of naturally occurring Drosophila.
|
|
Trustees of Leland Stanford University
|
|
Remarks
|
There is a PCT application
(WO
01/70816 A2) titled "Novel ecdysone receptor-based inducible gene
expression system", which was filed by Rohm and Hass Co. It
discloses gene expression modulation systems with two expression cassettes, one
having a DNA-binding and ligand-binding domain and in the other one a
transactivation domain and a ligand-binding domain. The ligand-binding domains
are derived from a retinoid X receptor, an ultraspiracle and an ecdysone
receptor. The DNA-binding domain is derived from GAL4 and LexA. Mutations to the
ecdysone or retinoid receptor enhance the ligand-binding activity and
non-steroid binding activity. Related applications are filed in the United
States
(US
2002/119521 A1 and
US
2004/96942 A1).
|
|
Note: Patent information on this page was last updated on 2 May 2006.
Metal-regulated promoters
Scientific aspects
Metallothioneins are proteins that bind
and sequester ionic forms of certain metals in fungi (yeast), plants and
animals. Such metals include copper, zinc, cadmium, mercury, gold, silver,
cobalt, nickel and bismuth. The specific metals sequestered by metallothioneins
vary in different organisms. Typically, these proteins are cysteine (cys)-rich
and lack aromatic amino acids. The presence of cys motifs is related to the
capability of binding metal ions. For instance, Cys-Xaa-Cys motifs have been
implicated in the binding and sequestration of copper. The location of these
residues within the protein is highly conserved between different species
showing a tight evolutionary relationship. In plants, metallothioneins have
roles in metal accumulation, metal detoxification and embryogenesis.
In the early 1990's, a gene expression system for plants was devised based on
the copper-dependent yeast metallothionein gene, named ACE1 (activating
copper-metallothionein expression). When a transcription factor-binding site for
the ACE1 gene was linked to the 35S CaMV promoter, the expression of
beta-glucuronidase (GUS) reporter gene was regulated in transformed tobacco
plants. Moreover, GUS expression depended on the presence of copper ions. The
yeast copper-inducible system has been also introduced in Arabidopsis
thaliana, driving the expression of green fluorescent protein (GFP).
Several metallothionein genes have been isolated from plants such as pea,
maize, barley, soybean and Arabidopsis and different plant tissues such
as maize root. Although the metal-responsive system is useful for regulation of
gene expression in a particular tissue and development stage, effects that the
metal could have in other tissues in the transformed organisms should be
assessed.
IP issues
A eukaryotic metal-responsive promoter, in general, was claimed in the
European patent
EP
94428, granted to University Patents Inc. (now
Competitive Technologies Inc.) on February 12, 1992. The
invention consisted of processes for securing the transcription of a selected
gene sequence in a host cell under the regulation of a eukaryotic
metal-responsive promoter. This European patent expired in all
European states where the patent was converted. The disclosed invention is now
in the public domain.
Other patents relevant to metal-responsive promoters are directed to
metallothionein gene promoters isolated from different sources such as yeast,
mouse, human and maize. In general, the patents claim:
- isolated sequences of metallothionein promoters;
- vectors for transformation of eukaryotes comprising the isolated genes; and
- methods for controlling the transcription of a gene of interest by the use
of metallothionein promoters.
The use of particular plant metallothionein promoters for transcription of
genes in a tissue of preference is the subject of several patents such as the
United States patent
US
6410828 and PCT patent application
WO
00/537763 by Dow Agrosciences. An isolated DNA sequence of a maize
metallothionein promoter having embryo-specific and root-specific expression is
disclosed in each document, respectively. Expression vectors containing the
metallothionein promoter and methods for expressing a heterologous gene sequence
in a transformed plant are also part of the inventions. Whether the promoters
are inducible by metals is not part of the disclosed. However, use of the maize
metallothionein promoter regardless of inducibility would be covered by the
claims.
Patents on Yeast Copper-Responsive Promoter
Genentech has a granted United States patent related to inducible
transcription control sequences isolated from a yeast metallothionein gene.
These transcriptional control sequences are also known as yeast copper chelatin
promoter.
The transcriptional control sequences of the invention include the
transcriptional promoter and the metal ion regulatory region. As well, in a
vector to be replicated in suitable host, a desired gene is under the control of
a yeast chelatin promoter, a yeast chelatin transcriptional control sequence, or
a metal ion regulatory region from yeast chelatin.
The yeast chelatin transcriptional control sequence contains the chelatin
promoter, including transcription initiation and ribosome binding sites. This is
a useful promoter in its own right, but to be induced by metal ions, the metal
ion regulatory region is needed. This last region is also useful when ligated to
other, non-metallothionein promoters, which are recognized by the desired host.
An isolated DNA sequence of a yeast chelatin metal ion regulatory region is
also part of the protected invention.
|
Patent Number
|
Title, Independent Claims and Summary of Claims
|
Assignee
|
|
US
4940661
- Earliest priority - 17 May 1984
- Filed - 29 September 1987
- Granted 10 July 1990
- Expected expiry - 29 September 2007
|
Title - Metallothionein transcription control sequences and
use thereof
Claim 1
A vector comprising a selection gene, yeast origin of
replication or autonomously replicating sequence and DNA encoding a eukaryotic
polypeptide other than yeast chelatin, said DNA encoding a eukaryotic
polypeptide that is under the control of
(a) a yeast chelatin promoter,
(b) a yeast chelatin transcription control sequence, or
(c) a metal ion regulatory region of the yeast chelatin transcription
control sequence which is free of the yeast chelatin promoter, whereby the
vector is replicable in a suitable host.
|
Claim 14
An isolated DNA sequence comprising a yeast chelatin metal
ion regulatory region free of the yeast chelatin promoter and free of DNA
encoding yeast chelatin.
|
|
Genentech
Inc.
|
Note: Patent information was last updated on 19 May 2006. Search terms:
"Metallothionein" in abstract and "Genentech" in applicant. Patent database:
PatentLens.
Patent on a mouse metallothionein promoter
The granted United States patent of University Patents Inc.
(now Competitive Technologies Inc.) relates to the
metal-responsive promoter/regulator sequences of the mouse
metallothionein-I gene.
The mouse metallothionein-I gene promoter of the invention is operatively
linked to a DNA sequence of interest, which is expressed in response to
variations in the concentration of metal ions in the environment. The host
organism, which stably incorporates the DNA sequence of interest, is a mammalian
host cell.
|
Patent Number
|
Title, Independent Claims and Summary of Claims
|
Assignee
|
|
US
4579821
- Earliest priority - 23 November 1981
- Filed - 23 November 1981
- Granted - 1 April 1986
- Expired - 1 April 2003
|
Title - Control of DNA sequence transcription
Claim 1
A process for subjecting the transcription of a selected DNA sequence to
external control under given environmental conditions which
comprises the steps of:
providing a selected isolated structural gene that is transcriptionally
responsive to a mouse metallothionein-I promotor/regulator DNA sequence under
the given environmental conditions; and
operatively fusing the selected structural gene with said promoter/regulator
DNA sequence.
|
Claim 2
In the genetic engineering process for securing transcription and expression
of a selected isolated structural gene sequence in a mammalian host cell wherein
said selected structural gene is stably incorporated as a chromosomal or
extrachromosomal constituent of the host, the improvement
comprising the step of:
operatively fusing with said selected structural gene sequence a mouse
metallothionein-I promoter/regulator DNA sequence which is responsive to
environmental variations within the host cell in the concentration of ions of
metal.
|
Claim 3
A fusion gene product, suitable for use in genetic transformation of a
mammalian host cell, said product comprising: a non mouse
metallothionein structural gene sequence to be incorporated in said host cell
operatively fused with a mouse metallothionein-I promoter/regulator DNA
sequence.
|
|
University Patents Inc.
(now
Competitive
Technologies)
|
|
Remarks
|
The related granted European patent
EP
94428 lapsed in all European contracting states.
Related application filed in Japan (JP 7083715 B4).
|
Note: Patent information was last updated on 19 May 2006. Search terms:
"transcription" in abstract and "University patents" in applicant. Patent
database: PatentLens in combination with INPADOC.
Patent on a human metallothionein promoter
The mammalian metallothionein promoter system claimed by the
University of
California
(Berkeley) in a granted United States
patent relates to a human metallothionein II transcriptional
regulatory system, which allows for regulated control of expression by
heavy metals and glucocorticoids.
|
Patent Number
|
Title, Independent Claims and Summary of Claims
|
Assignee
|
|
US
4601978
- Earliest priority - 24 November 1982
- Filed - 24 November 1982
- Granted - 22 July 1986
- Expired - July 2003
|
Title - Mammalian metallothionein promoter system
Claim 1
A DNA sequence of less than 500 base pairs, said DNA sequence
comprising the human MT-II transcriptional regulatory system
further comprising the transcription initiation sequence.
|
Claim 5
A DNA construct capable of regulated expression of an inserted gene in a
mammalian host, said construct comprising an extrachromosomal
replication system recognized by a mammalian host, which replication system has
been joined to a human MT-II gene inducible regulatory system in vitro, said
construct having DNA sequence(s) defining at least one restriction site within
the transcriptional control of the regulatory region for insertion of the gene.
|
Claim 10
A DNA construct useful for expression of a structural gene to produce a
polypeptide in a mammalian host, said vector comprising
(a) a replicon from bovine papilloma virus and
(b) a regulatory system from a human MT-II gene comprising
a promoter and a terminator and having at least one restriction site downstream
from the promoter in the direction of transcription.
|
The claimed invention is drawn to:
- A DNA sequence encoding the human metallothionein II
transcriptional regulatory system, which includes the promoter region, the
transcriptional initiation sequence (CAP site) and the regulatory sequences
responsible for inducible transcription.
- A DNA construct where the human metallothionein II
transcriptional regulatory sequences are combined with an extra chromosomal
replication system that is recognized by a given mammalian host. The combined
system regulates the expression of an inserted gene in the mammalian host.
- A vector containing a replicon from a bovine papilloma virus and a human
metallothionein II transcriptional regulatory sequences and a
restriction site downstream of the regulatory sequences for the insertion of a
gene of interest to be expressed in a mammalian host. The bovine papilloma virus
replicon allows the vector to be replicated in the mammalian host.
|
University of
California (Berkeley)
|
|
Remarks
|
Another United states patent titled " High efficiency eukaryotic
metallothionein promoter system"
(US
4511652) also expired.
|
Note: Patent information was last updated on 19 May 2006. Search terms:
"metallothionein" in abstract and "University of California" in applicant.
Patent database: PatentLens.
Pathogenesis-related (PR) promoter
Scientific aspects
Pathogenesis-related (PR) proteins are a heterogeneous group of proteins
induced in plants by pathogen infection and exogenous chemicals. PR proteins
take part in the systemic acquired resistance (SAR) that
develops in a resistant plant upon infection with a pathogen. As these proteins
are responsive not only to pathogens but also to
chemicals, the promoter sequences of these proteins have become
attractive as inducible expression systems in plants. For this purpose, promoter
sequences from diverse PR proteins have been isolated from plants such as
Arabidopsis and maize. Recently, these systems have been developed for
human cells as well.
Chemicals such as salicylic acid,
ethylene, thiamine, benzol (1,2,3) thiadiazole-7-cabothionic acid S-methyl ester
(BTH) have been identified as inducers of PR proteins. One of the best studied
promoters is the PR-1a promoter from tobacco. The expression of the
beta-glucuronidase (gus) gene, (when driven by the PR-1a promoter),
increased 5-10 fold after 1-3 days of induction with salicylic acid. The same
chemical induced GUS expression levels 10-fold after 8 days of spraying in field
conditions. The PR-1a promoter has also been used to induce the expression of
Bacillus thuringienesis delta-endotoxin in transgenic plants.
Salicylic acid is a potent inducer but there are crop tolerance problems
associated with its use. In contrast, BTH does not have a phytotoxic effect and
has a longer-lasting induction response of the PR-1a promoter is achieved when
compared with salicylic acid. The compounds also differ in their mechanism of
action: salicylic acid induces expression only in the treated tissue, whereas
BTH moves systemically through the plant.
Some of the drawbacks of PR gene promoters are their inducibility by common
environmental stimuli such as UV-B, ozone, and also oxidative stress. This
feature might complicate the control of gene expression by PR promoters in
non-laboratory conditions.
IP issues
The patents covered in this section relate to DNA sequences of isolated
PR-plant gene promoters, methods for inducing the transcription of a gene of
interest in a plant under the control of a PR promoter, and methods for finding
inducers of PR promoters. Novartis and Pioneer
Hi-Bred are the main entities having patents related to these aspects.
Broad and in force patents or patent applications directed to
any PR promoter or its use as inducible promoters in
transformed cells were not found.
A large portfolio comprising two patent families filed by and granted to
Novartis cover:
- DNA sequences of PR promoters isolated from Arabidopsis, tobacco
and cucumber;
- methods for inducing the transcription of a gene of interest in a plant
under the control of a PR promoter; and
- methods of inducing specific tobacco or Arabidopsis PR promoters. The claims
of these patents are however, narrow because specific plant PR-1 promoters are
covered
Novartis' portfolio also includes patents granted on isolated PR proteins,
methods for isolating chemically-regulatable sequences and methods for screening
agrochemicals that have the ability to induce SAR response. Because these areas
are outside the scope of this paper, they are not included in the present
analysis.
Pioneer's United States granted patent and European patent
application are directed to isolated DNA sequences of maize PR-1 gene promoters,
methods of using the promoters, and transformed plants. The isolated promoters
include both inducible and constitutive promoters. Some of the stimuli that
induce the maize PR-1 promoters are pathogen invasion, externally applied
chemicals, and environmental stresses. Plant cells and stably transformed plants
containing the promoters are also part of the invention.
See table next page for summary of the related patents.
The following table presents bibliographic information on the patents and
patent applications, and independent claims and summary of the claimed
inventions. It should be noted that Novartis and Pioneer Hi-bred patents are now
most likely controlled by Syngenta and DuPont, respectively. The contact
information is:
Syngenta: Intellectual Property Department, Schwarzwaldallee 215, CH-4058 Basel;
http://www.tmri.org/en/partnership/tech_introduction.aspx.
DuPont:
www.dupont.com/corp/contactus.html.
|
Patent/application number
|
Title, Independent claims and Summary of the claims
|
Assignee
|
|
US
5654414
- Earliest priority - 19 May 1995
- Filed - 19 May 1995
- Granted - 5 August 1997
- Expected expiry - 19 May 2015
|
Title - Chemically inducible promoter of a cucumber
chitinase/lysozyme gene
Claim 1
A nucleic acid promoter fragment isolated from the 5' flanking region
upstream of the coding region of a cucumber chitinase/lysozyme gene that is
inducible by application of benzo-1,2,3-thiadiazoles.
|
This patent mainly claims for an isolated promoter region of a
cucumber chitinase/lysozyme gene that is inducible by
application of benzo 1,2,3,-thiadiazoles (BTH). Chitinases are
pathogenesis-related enzymes induced in the SAR response.
|
Novartis Finance Corporation
|
|
US
5689044
- Earliest priority - 19 May 1995
- Filed - 24 May 1995
- Granted - 18 November 1997
- Expected expiry - 19 May 2015
|
Title - Chemically inducible promoter of a plant PR-1 gene
|
Claim 1
A chemically inducible nucleic acid promoter fragment isolated from the 5'
flanking region upstream of the coding region of a tobacco
PR-1a gene, wherein said promoter fragment comprises a
nucleotide fragment of at least 603-bp adjacent to the coding region of said
tobacco PR-1a gene, wherein said promoter fragment is inducible by application
of a benzo-1,2,3-thiadiazole, an isonicotinic acid compound, or a salicylic acid
compound.
|
|
Claim 3
A chemically inducible nucleic acid promoter fragment isolated from the 5'
flanking region upstream of the coding region of an
Arabidopsis PR-1 gene, wherein the coding region of
said Arabidopsis PR-1 gene comprises
the DNA sequence set forth in
SEQ
ID NO:33 or a DNA sequence which would encode the protein encoded by
SEQ
ID NO:33, wherein said promoter fragment is inducible by application of a
benzo-1,2,3-thiadiazole, an isonicotinic acid compound, or a salicylic acid
compound.
|
The claiming elements are:
- An isolated region of at least 603 bp that constitutes the tobacco
PR-1a gene promoter. The promoter is inducible by application of
salicylic acid, BTH and 2,6-dichloroisonicotinic acid (INA).
- The isolated promoter region of an
Arabidopsis PR-1 gene. This promoter is inducible by
the same compounds as the tobacco PR-1a gene promoter.
|
|
US
5789214
- Earliest priority - 19 May 1995
- Filed - 31 May 1995
- Granted - 4 August 1998
- Expected expiry - 4 August 2015
|
Title - Method of inducing gene transcription in a plant
|
Claim 1
A method of inducing gene transcription in a plant or plant tissue,
comprising the steps of:
(a) transforming said plant or plant tissue, each with a chimeric gene
comprising:
(i) a chemically inducible nucleic acid promoter fragment of at least
603-bp isolated from the 5' flanking region adjacent the coding region of a
tobacco PR-1a gene, and
(ii) a coding sequence of interest operatively linked to said promoter
fragment; and
(b) exposing said transgenic plant or plant tissue to a
benzo-1,2,3-thiadiazole, an isonicotinic acid compound, or a salicyclic acid
compound, whereby transcription of said coding sequence of interest is induced
in said plant or plant tissue.
|
|
Claim 8
A method of inducing gene transcription in a plant or plant tissue,
comprising the steps of:
(a) transforming said plant or plant tissue, each with a chimeric gene
comprising:
(i) a chemically inducible nucleic acid promoter fragment isolated from
the 5' flanking region adjacent the coding region of an Arabidopsis PR-1 gene,
wherein said Arabidopsis PR-1 gene comprises a DNA sequence that specifically
hybridizes
to SEQ
ID NO:33 or wherein said Arabidopsis PR-1 gene comprises a DNA sequence that
encodes the protein encoded by
SEQ
ID NO:33, and
(ii) a coding sequence of interest operatively linked to said promoter
fragment; and
(b) exposing said transgenic plant or plant tissue to a
benzo-1,2,3-thiadiazole, an isonicotinic acid compound, or a salicylic acid
compound, whereby transcription of said coding sequence of interest is induced
in said plant or plant tissue.
|
The claims are mainly to:
- Methods for inducing gene transcription in a plant or a plant tissue by
transforming such plant with a chimeric gene comprising the tobacco
PR-1a gene promoter or the Arabidopsis PR-1
gene promoter linked to a gene of interest and
applying an inducer to the transformed plant. The inducers are selected from
salicylic acid, BTH and isonicotinic acid (INA).
|
|
AU
708850 B2
- Earliest priority - 23 July 1995
- Filed - 18 July 1997
- Granted - 12 August 1999
- Expected expiry - 18 July 2017
|
Title - Chemically-inducible Arabidopsis PR-1 promoter
|
Claim 1
An isolated DNA molecule comprising a nucleotide sequence
selected from the following group:
a) a full-length chemically inducible promoter fragment
comprising nucleotides 1 through 4258 of SEQ ID NO:1;
b) an 81 5-bp long chemically inducible promoter fragment
comprising nucleotides 3444 through 4258 of SEQ ID NO:1;
and
c) a 698-bp long chemically inducible promoter fragment
comprising nucleotides 3561 through 4258 of SEQ ID NO:1.
|
|
Claim 14
An isolated DNA molecule involved in inducibility of a chemically inducible
promoter selected from the following group:
a) LS4 comprising nucleotides 3584 through 3593 of SEQ ID NO:1;
b) L57 comprising nucleotides 3614 through 3623 of SEQ ID NO:1;
c) LS 10 comprising nucleotides 3644 through 3653 of SEQ ID NO:1; and
d) a region spanning LS7-LSIO and comprising nucleotides 3614 through 3653
of SEQ ID NO:1.
|
The claiming elements are:
- Isolated full-length chemically-inducible Arabidopsis PR-1
promoter and isolated shorter portions of the promoter that are
required for induction of gene expression by chemicals such as salicylic acid,
BTH and 2,6-dichloroisonicotinic acid (INA).
- Isolated motifs in the promoter that when mutated alters the inducible
activity of the promoter
|
|
Remarks
|
Related applications also filed in Canada (CA 2232741 AA)
and Japan (JP 11513897 T2). The claims as filed of the Canadian
application are the same as the claims of the Australian granted patent. The
related European application EP 868 426 A1 was withdrawn on
January 2, 2003.
|
|
US
6429362
- Earliest priority - 26 February 1998
- Filed - 25 February 1999
- Granted - 6 August 2002
- Expected expiry - 25 February 2019
|
Title - Maize PR-1 gene promoters
|
Claim 1
An isolated promoter comprising a nucleotide sequence that initiates
transcription in a plant cell, wherein said nucleotide sequence is selected from
the group consisting of:
a) a nucleotide sequence comprising the sequence set forth in
SEQ
ID NO: 3 or
4;
and
b) a nucleotide sequence comprising the plant promoter sequence deposited in
the plasmid designated as ATCC Accession No. 207139 or 207131.
|
|
Claim 5
A method for driving expression of a heterologous nucleotide sequence in a
plant, said method comprising the steps of:
a) transforming a plant cell with an expression cassette comprising the
heterologous nucleotide sequence operably linked to a promoter that initiates
transcription in a plant cell, wherein said promoter is selected from the group
consisting of:
i) a nucleotide sequence comprising the sequence set forth in
SEQ
ID NO: 3 or
4;
and
ii) a nucleotide sequence comprising the plant promoter sequence
deposited in the plasmid designated as ATCC Accession No. 207139 or 207131; and
b) regenerating a stably transformed plant from said plant cell.
|
|
Claim 9
A plant cell transformed with a DNA construct comprising a heterologous
nucleotide sequence operably linked to a promoter that initiates transcription
in said plant cell, wherein said promoter comprises a nucleotide sequence
selected from the group consisting of:
a) a nucleotide sequence comprising the sequence set forth in
SEQ
ID NO: 3 or
4;
and
b) a nucleotide sequence comprising the plant promoter sequence deposited in
the plasmid designated as ATCC Accession No. 207139 or 207131.
|
|
Claim 13
A plant stably transformed with a DNA construct comprising a heterologous
nucleotide sequence operably linked to a promoter that initiates transcription
in a plant cell, wherein said promoter comprises a nucleotide sequence selected
from the group consisting of:
a) a nucleotide sequence comprising the sequence set forth in
SEQ
ID NO: 3 or
4;
and
b) a nucleotide sequence comprising the plant promoter sequence deposited in
the plasmid designated as ATCC Accession No. 207139 or 207131.
|
The claims are generally drawn to:
- Two isolated DNA sequences of inducible maize PR-1 gene
promoters
- A method to drive the expression of a heterologous gene in a plant by using
the claimed maize PR-1 promoters
- A plant cell transformed with a DNA construct comprising either of the two
promoters, and
- A plant stably transformed with such a construct
|
Pioneer Hi-Bred International Inc.
|
|
AU
754376 B2
- Earliest priority - 26 February 1998
- Filed - 11 February 1999
- Granted - 14 November 2002
- Expected expiry - 11 February 2019
|
Title - Family of maize PR-1 genes and promoters
|
Claim 1
An isolated nucleic acid molecule having a nucleotide sequence for a promoter
that is capable of initiating transcription in a plant cell, wherein said
nucleotide sequence is selected from the group consisting of:
a) a nucleotide sequence comprising the sequence set forth in SEQ ID NO:
1,2,3,4, or 5;
b) a nucleotide sequence comprising at least 40 contiguous nucleotides of
the sequence set forth in SEQ ID NO: 1,2,3,4, or 5; and
c) a nucleotide sequence that hybridizes under stringent conditions to a
sequence of a) or b).
|
|
Claim 5
A method for inducing expression of a heterologous nucleotide sequence in a
plant, said method comprising transforming a plant cell with a DNA construct
comprising said heterologous nucleotide sequence operably linked to a promoter
that is capable of initiating transcription in a plant cell in response to a
stimulus, regenerating a stably transformed plant from said plant cell, and
exposing said plant to said stimulus, wherein said promoter comprises a
nucleotide sequence selected from the group consisting of:
a) a nucleotide sequence comprising the sequence set forth in SEQ ID
NO:1,2,3, or 4;
b) a nucleotide sequence comprising at least 40 contiguous nucleotides of
the sequence set forth in SEQ ID NO: 1,2,3, or 4; and
c) a nucleotide sequence that hybridizes under stringent conditions to a
sequence of a) or b).
|
|
Claim 9
A method for constitutively expressing a heterologous nucleotide sequence in a
plant, said method comprising transforming a plant cell with a DNA construct
comprising said heterologous nucleotide sequence operably linked to a promoter
that is capable of initiating constitutive transcription in a plant cell and
regenerating a stably transformed plant from said plant cell, wherein said
promoter comprises a nucleotide sequence selected from the group consisting of:
a) a nucleotide sequence comprising the sequence set forth in SEQ ID NO: 5;
b) a nucleotide sequence comprising at least 40 contiguous nucleotides of
the sequence set forth in SEQ ID NO:5; and
c) a nucleotide sequence that hybridizes under stringent conditions to a
sequence of a) or b).
|
|
Claim 13
A plant cell stably transformed with a DNA construct comprising a heterologous
nucleotide sequence operably linked to a promoter that is capable of initiating
transcription in said plant cell, wherein said promoter comprises a nucleotide
sequence selected from the group consisting of:
a) a nucleotide sequence comprising the sequence set forth in SEQ ID NO:
1,2,3,4, or 5;
b) a nucleotide sequence comprising at least 40 contiguous nucleotides of
the sequence set forth in SEQ ID NO: 1,2,3,4, or 5; and
c) a nucleotide sequence that hybridizes under stringent conditions to a
sequence of a) or b).
|
|
Claim 17
A plant stably transformed with a DNA construct comprising a heterologous
nucleotide sequence operably linked to a promoter that is capable of initiating
transcription in a plant cell, wherein said promoter comprises a nucleotide
sequence selected from the group consisting of:
a) a nucleotide sequence comprising the sequence set forth in SEQ ID NO:
1,2,3,4, or 5;
b) a nucleotide sequence comprising at least 40 contiguous nucleotides of
the sequence set forth in SEQ ID NO: 1,2,3,4, or 5; and
c) a nucleotide sequence that hybridizes under stringent conditions to a
sequence of a) or b).
|
|
Claim 22
An isolated nucleic acid molecule having a nucleotide sequence selected from the
group consisting of:
a) a nucleotide sequence comprising the sequence set forth in SEQ ID NO:
6,8,10, or 14;
b) a nucleotide sequence encoding a polypeptide comprising the amino acid
sequence set forth in SEQ ID NO: 7,9,11, or 15;
c) a nucleotide sequence that shares at least 85% sequence identity to the
sequence set forth in SEQ ID NO:6, 8 or 14; and
d) a nucleotide sequence that shares at least 90% sequence identity to the
sequence set forth in SEQ ID NO:10.
|
|
Claim 26
A method for creating or enhancing disease resistance in a plant, said method
comprising transforming said plant with a DNA construct comprising a PR-1
sequence operably linked to a promoter that drives expression of a coding
sequence in a plant cell and regenerating stably transformed plants, wherein
said PR-1 sequence is selected from the group consisting of:
a) a nucleotide sequence comprising the sequence set fortha in SEQ ID NO: 6,
8, 10, or 14;
b) a nucleotide sequence encoding a polypeptide comprising the amino acid
sequence set forth in SEQ ID NO: 7, 9, 11, or 15;
c) a nucleotide sequence comprising at lease 16 contiguous nucleotides of a
sequence of a) or b); and
d) a nucleotide sequence that hybridizes under stringent conditions to a
sequence of a), b), or c).
|
|
Claim 33
A plant cell stably transformed with a DNA construct comprising a PR-1 sequence
operably linked to a promoter that drives expression of a coding sequence plant
cell, wherein said PR-1 sequence is selected from the group consisting of:
a) a nucleotide sequence comprising the sequence set forth in SEQ ID NO: 6,
8, 10 or 14;
b) a nucleotide sequence encoding a polypeptide comprising the amino acid
sequence set forth in SEQ ID NO:7, 9, 11 or 15;
c) a nucleotide sequence comprising at least 16 contiguous nucleotides of a
sequence of a) or b); and
d) a nucleotide sequence that hybridizes under stringent conditions to a
sequence of a, b) or c).
|
Claim 34
A plant stably transformed with a DNA construct comprising a PR-1 sequence
operably linked to a promoter that drives expression of a coding sequence in a
plant cell, wherein said PR-1 sequence is selected from the group consisting of:
a) a nucleotide sequence comprising the sequence set forth in SEQ ID NO: 6,
8, 10, or 14;
b) a nucleotide sequence encoding a polypeptide comprising the amino acid
sequence set forth in SEQ ID NO: 7, 9, 11 or 15;
c) a nucleotide sequence comprising at least 16 contiguous nucleotides of a
sequence of a) or b); and
d) a nucleotide sequence that hybridizes under stringent conditions to a
sequence of a), b) or c).
|
The claims include:
- DNA sequences that hybridize under stringent conditions to the promoter
sequences;
- a DNA sequence of a constitutive maize PR-1 gene promoter;
- methods for inducing the expression of a gene of interest or constitutively
expressing the gene of interest according to the promoter used;
- the nucleotide sequences of the maize PR-1 genes;
- methods for enhancing disease resistance in a plant by using the claimed
PR-1 sequences; and
- plant cells and stably transformed plants with such PR-1 genes.
|
|
Remarks
|
Related application in Europe (EP 1056862) has been
withdrawn and the application in Canada (CA 2315549 AA) is also
dead.
|
Note: Patent information on this page was last updated on 2 May 2006.
Search terms for Novartis: "transcription" in abstract and "Novartis" in
applicant.
Search terms for Pioneer Hi-Bred: "promoters" in abstract and "Pioneer
Hi-Bred" in applicant. Patent database: PatentLens in combination with INPADOC.
Physically-regulated promoters
Summary
Promoters induced by environmental
factors such as water or salt stress, anaerobiosis, temperature, illumination
and wounding have potential for use in the development of plants resistant to
various stress conditions. These promoters contain regulatory elements that
respond to such environmental stimuli.
Temperature-induced promoters include cold- and heat-shock-induced promoters.
In many cases, these promoters are able to operate under normal temperature
conditions, which vary according to the organism, but when either cold or heat
is applied, the promoters maintain activity. In addition, expression can be
enhanced by the application of higher or lower temperature as compared to the
normal temperature conditions. One of the best studied eukaryotic heat-shock
systems is the one found in Drosophila (fruit fly).
In plants, light-regulated promoters are critical in regulating plant growth
and development through the modulation of expression of light-responsive genes.
Light-responsive elements from genes such as the small subunit of
ribulose-1,5-bisphosphate carboxylase-oxygenase (rbcS) gene, the chlorophyl a/b
binding protein, and the chalcone synthase have been widely studied. A molecular
dissection of their sequences has shown multiple cis-acting elements for
light-dependent gene expression. The light-regulated elements in these promoters
exhibit some properties of enhancer-like elements. It is unlikely that a single
cis element is sufficient to confer light responsiveness to a non-light
regulated promoter. In addition, light-regulated gene expression is under the
influence of multiple environmental factors.
The patents presented in this section relate to promoters responsive to
temperature and light. The promoters are
derived from genes from multiple organisms including bacteria and plants.
Methods to use promoters responsive to heat, cold, light or darkness in general
terms are part of the inventions discussed here. In some cases, such as in
patents related to promoters responsive to cold and light stimuli, particular
DNA sequences of promoters are claimed.
The selected analyzed patents are categorized as follows:
Temperature-regulated promoters
IP issues
1. Heat-inducible promoters
Mycogen Plant Sciences, The United States Department of Health and Human
Services and The General Hospital Corporation have granted patents and patent
applications that relate in general to DNA sequences of heat shock promoters and
methods for expressing a gene of interest under the control of such promoters.
Some of the inventions relate to the use of the heat shock promoters in
transformed plants, while others do not specify the organism to be transformed.
For some properties of heat shock promoters please also refer to
the plant
ubiquitin promoters.
A. Mycogen Plant Sciences' patents
The patents granted to Mycogen in the United States, Europe and Canada are
directed to:
- consensus DNA sequences of plant heat shock promoters;
- bacterial strains containing a vector with a consensus heat shock promoter
and a gene of interest under its control for expression in plants;
- recombinant Agrobacterium Ti-plasmids comprising a heat shock
promoter;
- methods for expressing a heterologous gene under the control of a heat shock
promoter in plant cells; and
- methods for identifying transformed plants.
patent claims in particular the use of a soybean heat shock promoter
to drive the expression of a structural gene in a plant. The soybean
heat shock promoter and the structural gene are engineered in a plasmid carried
by an Agrobacterium strain used to infect the plant to be transformed.
Mycogen Plant Sciences has changed its name to Mycogen Seeds and is now an
affiliate of Dow AgroSciences LLC. Therefore , Mycogen patents and patent
applications are probably controlled by Dow AgroSciences LLC. The contact
information can be found at http://www.dowagro.com/contact/index.htm or
http://www.dowagro.com/mycogen/contact/index.htm.
|
Patent Number
|
Title, Independent Claims and Summary of Claims
|
Assignee
|
|
US
5447858
- Earliest priority - 5 September 1984
- Filed - 5 September 1984
- Granted - 5 September 1995
- Expected expiry - 5 September 2012
|
Title - Heat shock promoter and gene
Claim 1
A recombinant heat shock gene comprising a plant heat shock
promoter and a heterologous structural gene whose expression is controllable
thereby, wherein said promoter comprises the consensus
nucleotide sequence 5'-C-T-X-G-A-A-X-X-T-A-C-X-X-X-3', where X is A,T,C or G.
|
Claim 2
A recombinant heat shock gene comprising a plant heat shock
promoter and a heterologous structural gene whose expression is controlled
thereby, wherein said promoter comprises the consensus
nucleotide sequence 5'-C-T-S-G-A-A-M-R-T-A-C-W-M-K-3' where S is C or G; M is A
or C; R is A or G; W is A or T; and K is T or G.
|
Claim 8
A vector comprising a fragment of DNA capable of functioning
as a plant heatshock promoter wherein said fragment of DNA
comprises the consensus nucleotide sequence
5'-C-T-X-G-A-A-X-X-T-A-C-X-X-X-3', where X is A, T, C cr G.
|
Claim 9
A vector comprising a fragment of DNA capable of functioning
as a plant heat shock promoter wherein said fragment comprises
the consensus nucleotide sequence of 5'-C-T-S-G-A-A-M-R-T-A-C-W-M-K-3' where S
is C or G; M is A or C; R is A or G; W is A or T; and K is T or G.
|
Claim 19
A bacterial strain containing therein recombinant DNA
comprising:
(a) a fragment of DNA capable of
controlling heatshock expression of a gene in a plant; and
(b) a
structural gene oriented with respect to said fragment of DNA so as to be
expressible under the control thereof wherein said fragment of DNA capable of
controlling heat shock expression of a gene in a plant
comprises the consensus nucleotide sequence
5'-C-T-X-G-A-A-X-X-T-A-C-X-X-X-3', where X is A, T, C or G.
|
Claim 20
A bacterial strain containing therein recombinant DNA
comprising:
(a) a fragment of DNA capable of
controlling heat shock expression of a gene in a plant; and
(b) a
structural gene oriented with respect to said fragment of DNA so as to be
expressible under the control thereof wherein the fragment of DNA capable of
controlling heat shock expression of a gene in a plant
comprises the consensus nucleotide sequence
5'-C-T-S-G-A-A-M-R-T-A-C-W-M-K-3' where S is C or G; M is A or C; R is A or G; W
is A or T; and K is T or G.
|
Claim 34
A method for recognizing a plant cell containing recombinant DNA
comprising the steps of:
(1) transferring into said
plant cell recombinant DNA comprising:
(a) a heat shock promoter
which comprises the consensus sequence 5'-C-T-X-G-A-A-X-X-T-A-C-X-X-X-3', where
X is A, T, C, or G;
(b) a transformation recognition gene under
the control of said heat shock promoter capable of causing an observable or
detectable reaction when expressed;
(2) applying a stress to said
plant cell in which it is desired to recognize those containing recombinant DNA,
which stress is capable of inducing a response in said heat shock promoter; and
(3) observing or detecting the reaction caused by expression of said
transformation recognition gene under the control of said heat shock promoter to
recognize said plant cell containing recombinant DNA including said
transformation recognition gene.
|
Claim 36
A method for recognizing a plant cell containing recombinant DNA
comprising the steps of:
(1) transferring into said
plant cell recombinant DNA comprising:
(a) a heat
shock promoter which comprises the consensus sequence
5'-C-T-S-G-A-A-M-R-T-A-C-W-M-K-3' where S is C or G; M is A or C; R is A or G; W
is A or T; and K is T or G;
(b) a transformation recognition gene
under the control of said heat shock promoter capable of causing an observable
or detectable reaction when expressed;
(2) applying a stress to said
plant cell in which it is desired to recognize those containing recombinant DNA,
which stress is capable of inducing a response in said heat shock promoter; and
(3) observing or detecting the reaction caused by expression of said
transformation recognition gene under the control of said heat shock promoter to
recognize said plant cell containing recombinant DNA including said
transformation recognition gene.
|
|
Mycogen Plant Sciences
|
|
EP
159884 B1
- Earliest priority - 13 April 1984
- Filed - 12 April 1985
- Granted - 10 February 1993
- Expected expiry - 12 April 2005
|
Title - Heat shock promoter and gene
The claims are the same as
US
5447858 (see above).
|
CA 1338010
- Earliest priority - 13 April 1984
- Filed - 11 April 1985
- Granted - 30 January 1996
- Expected expiry - 29 January 2013
|
Title - Heat shock promoter and gene
This Canadian patent claims in particular the use of a soybean heat
shock promoter to drive the expression of a structural gene in a plant.
The soybean heat shock promoter and the structural gene are engineered in a
plasmid carried by an Agrobacterium strain used to infect the plant to
be transformed.
Claim 1
A recombinant heat shock gene comprising a plant heat shock
promoter and a heterologous structural gene whose expression is controllable
thereby, wherein said promoter comprises the consensus
nucleotide sequence 5'-C-T-X-G-A-A-X-X-T-A-C-X-X-X-3', where X is A,T,C or G.
|
Claim 8
A vector comprising a fragment of DNA capable of functioning
as a plant heatshock promoter wherein said fragment of DNA
comprises the consensus nucleotide sequence
5'-C-T-X-G-A-A-X-X-T-A-C-X-X-X-3', where X is A, T, C cr G.
|
Claim 19
A bacterial strain containing therein recombinant DNA
comprising:
(a) a fragment of DNA capable of
controlling heatshock expression of a gene in a plant; and
(b) a
structural gene oriented with respect to said fragment of DNA so as to be
expressible under the control thereof wherein said fragment of DNA capable of
controlling heat shock expression of a gene in a plant
comprises the consensus nucleotide sequence
5'-C-T-X-G-A-A-X-X-T-A-C-X-X-X-3', where X is A, T, C or G.
|
Claim 34
A method for obtaining a transiet increase in expression level of a
heterologous structural gene in a plant comprising
inserting into said genome a DNA fragment comprising a promoter
capable of inducing a heat shock reaponse in a plant exposed to stress, which
promoter comprises the consensus nucleotide sequence
5'-C-T-X-G-A-A-X-X-T-A-C-X-X-X-3', where X is A, T, C, or G, said promoter being
combined with said structural gene such that said structural gene is expressed
under the control of said promoter, followed by
exposing said plant to
stress such that said promoter responds thereby obtaining a transient increase
in expression level of said structural gene in said plant.
|
|
Claim 54
A method for recognizing a plant cell containing recombinant DNA
comprising the steps of:
(1) transferring into said
plant cell recombinant DNA comprising:
(a) a heat
shock promoter which comprises the consensus sequence
5'-C-T-X-G-A-A-X-X-T-A-C-X-X-X-3', where X is A, T, C, or G;
(b) a
transformation recognition gene under the control of said heat shock promoter
capable of causing an observable or detectable reaction when expressed;
(2) applying a stress to said plant cell in which it is desired to recognize
those containing recombinant DNA, which stress is capable of inducing a response
in said heat shock promoter; and
(3) observing or detecting the
reaction caused by expression of said transformation recognition gene under the
control of said heat shock promoter to recognize said plant cell containing
recombinant DNA including said transformation recognition gene.
|
|
|
Remarks
|
There are two related applications pending in Japan (JP 60248176
A2 and JP 7075567 A2).
|
Note: Patent information on this page was last updated on 4 May 2006.
Search terms: "Heat shock promoter" in abstract; "Mycogen Plant Sciences" in
applicant
Patent database: Patent Lens
B. The United States Department of Health and Human
Services' patents
The granted Australian patent and the European
application are related to methods for the controlled expression in a
gene of interest by a heat-inducible promoter in a spatial and
temporal fashion.
One of the methods of the invention is not directed to a
particular organism or cell, but to a cell mass or multicellular
organism. In another method, animal cells are selected
for the expression of a therapeutic protein under the control
of a heat shock promoter.
|
Patent Number
|
Title, Independent Claims and Summary of Claims
|
Assignee
|
|
AU
732872 B2
- Earliest priority - 15 August 1996
- Filed - 14 August 1997
- Granted - 3 May 2001
- Expected expiry - 14 August 2017
|
Title - Spatial and temporal control of gene expression
using a heat shock protein promoter in combination with local heat
Claim 1
A method for the spatial and temporal control of the expression of a gene of
interest within a preselected discrete region of a cell mass or multicellular
organism, comprising:
selectively heating a preselected discrete region of a cell mass or
multicellular organism that includes cells that contain a genetically engineered
gene of interest operably linked to a heat-inducible promoter, thereby inducing
the expression of said gene of interest in the cells that are selectively
heated.
|
Claim 9
A method of providing a therapeutic protein to selected cells in an animal,
comprising the steps of:
introducing into cells of an animal a DNA molecule having
a heat shock promoter sequence operably linked to and exerting regulatory
control over a sequence encoding a therapeutic protein, and
activating said heat shock promoter sequence through the application of a
focused ultrasound so that said DNA segment expresses a therapeutically
effective amount of said therapeutic protein.
|
|
The United States Department of Health and Human
Services
|
|
EP 922110 A2
- Earliest priority - 15 August 1996
- Filed - 14 August 1997
- Granted - Pending
- Expected expiry - N/A
|
The title and the claims are the same as AU 732872.
|
|
US
2002/165191
- Earliest priority - 15 August 1996
- Filed - 11 March 2002
- Granted - Pending
- Expected expiry - N/A
|
The title and the claims are the same as AU 732872.
|
Claim 1
A method for the spatial and temporal control of the expression of a gene of
interest within a preselected discrete region of a cell mass or multicellular
organism, comprising: selectively heating a preselected discrete region of a
cell mass or multicellular organism that includes cells that contain a
genetically engineered gene of interest operably linked to a heat-inducible
promoter, thereby inducing the expression of said gene of interest in the cells
that are selectively heated.
|
|
Claim 9
A method of providing a therapeutic protein to selected cells in an animal,
comprising the steps of: introducing into cells of an animal a DNA molecule
having a heat shock promoter sequence operably linked to and exerting regulatory
control over a sequence encoding a therapeutic protein, and activating said heat
shock promoter sequence through the application of a focused ultrasound so that
said DNA segment expresses a therapeutically effective amount of said
therapeutic protein.
|
|
|
US
2005/059623
- Earliest priority - 15 August 1996
- Filed - 9 June 2004
- Granted - Pending
- Expected expiry - N/A
|
This is a countinuation of
US
2002/165191 (see above) with claims 1-9 canceled
|
Claim 10
A method for the spatial and temporal control of the expression of a
genetically engineered gene of interest operably linked to a heat shock promoter
in host cells within a preselected region of a mammal, consisting: selectively
heating the region to non-lethal supraphysiological temperatures for a period of
time by applying electromagnetic radiation to the region, thereby spatially and
temporally controlling the expression of the gene of interest.
|
|
Claim 18
A method of treating mammal, the method comprising: introducing a genetically
engineered gene operably linked to a heat shock promoter into host cells of a
region of the mammal; selectively heating the region to non-lethal
supraphysiological temperatures by applying electromagnetic radiation for a
period of time to the region, thereby spatially and temporally controlling the
expression of the gene and amount of the protein produced in the region.
|
|
|
Remarks
|
Related application also filed in Japan (JP 2001501458 T2).
|
Note: Patent information on this page was last updated on 4 May 2006.
Search terms: "Heat shock promoter" in abstract; "the Department of Health
and Human Services" in applicant
Patent database: Patent Lens and esp@cenet
C. The General Hospital Corporation's
patents
The General Hospital Corporation has granted patents in Canada and Australia
and filed an application in Europe. However, this patent family was obviously
abandoned.
The disclosed inventions are mainly related to:
-
A method for the production of a polypeptide or protein whose structural gene
is under the control of an inducible heat shock promoter.
The structural gene and the heat inducible promoter are transformed in a host
cell, which is not limited to a particular host organism.
A gene amplification system under the control of a constitutive promoter is used
to increase the copy number of the structural gene.
-
A method for increasing the production of a protein in a dihydrofolate
reductase deficient mammalian cell. The structural gene of the protein of
interest is placed under the control of a heat shock promoter and a
dihydrofolate reductase gene is under the control of a constitutive promoter.
-
A host cell co-transformed with a gene amplification system as mentioned
before and a structural gene driven by an inducible heat shock promoter.
Dihydrofolate reductase is a small enzyme that plays a supporting role in the
building of DNA and other processes. It manages the concentration of folate, an
organic molecule that shuttles carbon atoms to enzymes that need them in their
reactions. Dihydrofolate reductase recycles folate after the release of its
carbon atoms.
The dihydrofolate reductase gene is used by the inventors as a gene
amplification system. Dihydrofolate reductase deficient mammalian cells
transformed with this gene are able to grow in the presence of methotrexate,
which is an inhibitor of dihydrofolate reductase. Growing in increasing
concentrations of methotrexate leads the cells to overproduce dihydrofolate
reductase. If a structural gene controlled by an inducible heat shock promoter
is co-transformed with the dihydrofolate reductase gene system, both genes are
amplified together under the selection pressure.
|
Title, Independent Claims and Summary of Claims
|
Assignee
|
|
CA 1324097
- Earliest priority - 6 February 1986
- Filed - 4 February 1987
- Granted -9 November 1993
- Lapsed - 11 May 1996
|
Title - Inducible heat shock and amplification system
Claim 1
A method for high level production of a polypeptide or protein in a
transformed host cell comprising:
(a) transforming a host cell with a structural gene encoding for a
polypeptide or protein, under the control of an inducible heat shock promoter;
(b) amplifying the copy number of said structural gene in said transformed
host cells using an amplification system urder the control of a promoter other
than an inducie e heat shock promoter;
(c) inducing said inducible promoter by heat shock to said transformedost
cells at a temperature and for a time sufficient to transcribe said structural
gene;;
(d) allowing said heat shocked cells to recover at a lower temperature than
said heat shock temperature and for a time sufficient to translate said
transcribed structural gene, producing said polypeptide or protein.
|
Claim 10
A method for increasing the production of a structural polypeptide or protein
in a transformed mammalian cell comprising the steps of:
(a) culturing a dihydrofolate reductase (DHFR) deficient mammalian cell
transformed with
(i) a DHFR gene under the control of a constitutive promoter. and
(ii) astructural gene encoding for a polypeptide or protein under the
control of an inducible heat shock promoter, in a culture medium containing
methotrexate, for a time sufficient to permit growth of said transformed
mammalian cells;
(b) inducing said heat shock promoter at a temperature and for a time
sufficient to transcribe said structural gene; and
(c) allowing said heat shocked cells to recover from said heat shock at a
lower temperature and for a time sufficient to translate said transcribed
structural gene, producing said polypeptide or protein.
|
Claim 17
A host cell cotransformed with:
(a) a gene amplification system under the control of a constitutive
promoter, and
(b) a structural gene encoding for a polypeptide or protein under control of
an inducible heat shock promoter.
|
|
General Hospital Corporation
|
|
Remarks
|
The related Australian granted patent AU 606049 B2, granted
on January 31, 1991, ceased on September 21, 1995. Similarly, the related
European patent application EP 232845 A2 was withdrawn on
October 23, 1991.
|
Note: Patent information on this page was last updated on 4 May 2006.
Search terms: "inducible heat shock" in abstract.
Patent database: Patent Lens and esp@cenet in combinaton with INPADOC.
2. Cold-inducible promoters
Promoters responsive to low or cold temperatures (equal to or below 10°C) are
the subject matter of patents and patent applications filed by the Japanese
companies Takara Shuzo Co., Ltd and Japan Tobacco Inc., The
University of Quebec
in Montreal and Danisco.
Most of the
cited institutes claim in their patents a particular "cold" promoter and its
variant sequences.
A. Takara Shuzo Co., Ltd's patents and patent applications
The United States granted patents and the European and Canadian patent
applications relate to a vector useful for the expression of a protein under low
temperature conditions. The vector of the invention contains:
- a promoter regulatory region; and
- a 5'-untranslated region derived from a cold-shock protein gene.
The promoter region and 5' untranslated region of the vector are not limited
to a particular promoter. In addition a promoter and a 5'-untranslated region
are derived from the cold-shock protein (csp) A of Escherichia coli.
The nucleotide sequence of such promoter is also claimed by the inventors.
|
Patent Number
|
Title, Independent Claims and Summary of Claims
|
Assignee
|
|
US
6479260
- Earliest priority - 20 November 1997
- Filed - 19 May 2000
- Granted - 12 November 2002
- Expected expiry - 19 May 2020
|
Title - Low-temperature inducible expression vector
Claim 1
A vector which is characterized in containing each of the following elements:
(1) a promoter which shows its action in the host to be used;
(2) regulatory region for regulating the action of the promoter of (1); and
(3) a region which codes for the 5'-untranslated region derived from
cold-shock protein gene mRNA or a region which codes for the region where
substitution, deletion, insertion or addition of at least one base is applied to
the said untranslated region and which contains a base sequence as shown in
SEQ
ID NO:1 in the Sequence Listing.
|
Claim 10
An isolated promoter consisting of a base sequence as shown in
SEQ
ID NO: 5 in the Sequence Listing.
|
Claim 11
An isolated promoter containing a base sequence as shown in
SEQ
ID NO: 5 in the Sequence Listing and consisting of a base sequence having
135 or less bases, wherein the promoter does not contain the region which is
ascribed to mRNA.
|
|
Takara Shuzo Co., Ltd
(now Takara Holdings Inc.)
|
|
US
6897042
- Earliest priority - 20 November 1997
- Filed - 10 October 2002
- Granted - 24 May 2005
- Expected expiry - 19 May 2020
|
Title - Low-temperature inducible expression vector
Claim 1
A vector which is characterized in containing each of the following elements:
(1) a promoter which is derived from cold-shock protein gene, and
which shows its action in the host to be used;
(2) regulatory region
for regulating the action of the promoter of (1), wherein said regulatory region
is located downstream of the promoter of (1); and
(3) a region which
codes for the 5'-untranslated region derived from cold-shock protein gene mRNA
or a region which codes for the region where substitution, deletion, insertion,
or addition of at least one base is applied to said untranslated region.
|
Claim 12
A method for the expression of the desired protein which is characterized in
containing the following steps:
1) a step where a host is transformed
with a vector containing:
a) a promoter which is derived from
cold-shock protein gene and which shows its action in the host to be used;
b) a regulatory region for regulating the action of the promoter
of a), wherein said regulatory region; and
c) a region which
codes for the 5'-untranslated region derived from cold-shock protein gene mRNA
or a region which codes for the region where substitution, deletion, insertion,
or addition of at least one base is applied to the untranslated region wherein a
gene coding for the desired protein is integrated;
2) a step where the
resulting transformant is incubated; and
3) a step where action of
promoter is induced via a function of a regulatory region and, at the same time,
incubating temperature is made lower than the ordinary temperature to express
the desired protein.
|
This patent is a Countinuation of
US
6479260.
|
|
EP 1033408 A1
- Earliest priority - 20 November 1997
- Filed - 17 November 1998
- Granted - Pending
- Expected expiry - N/A
|
Title - Cold-inducible expression vector
Claim 1
A vector which is characterized in containing each of the following elements:
(1) a promoter which shows its action in the host to be used;
(2)
regulatory region for regulating the action of the promoter of (1); and
(3) a region which codes for the 5'-untranslated region derived from cold-shock
protein gene mRNA or a region which codes for the region where substitution,
deletion, insertion or addition of at least one base is applied to the said
untranslated region.
|
Claim 14
A promoter containing a base sequence as shown in SEQ ID NO:5 in the Sequence
Listing and consisting of a base sequence having 135 or less bases.
|
|
|
CA 2309600 AA
- Earliest priority - 20 November 1997
- Filed - 17 November 1998
- Granted - Pending
- Expected expiry - N/A
|
Title - Cold-inducible expression vector
The claims of this Canadian patent application are the same as EP
1033408 (above).
|
|
Remarks
|
The related Australian application AU 10546/99 lapsed on
August 3, 2000.
|
Note: Patent information was last updated on 10 May 2006.
Search terms: "promoter" in abstract and "Takara Shuzo" in applicant.
Patent database: PatentLens.
B. Patents on particular cold-temperature promoters
As mentioned above, the patents filed by most institutes relate to a specific
low temperature-responsive promoter.
The following table summarizes some bibliographic data of the patents and
patent applications filed by the cited entities and a brief description of the
promoter claimed.
|
Patent number
|
Title, Independent Claims and Summary of Claims
|
Assignee
|
|
US
6084089
- Earliest priority - 27 December 1995
- Filed - 27 October 1997
- Granted - 4 July 2000
- Expected expiry - 27 October 2017
|
Title - Cold-inducible promoter sequences
Claim 1
A DNA sequence comprising a nucleotide sequence from the first to the 3546th
nucleotide in the nucleotide sequence shown in SEQ ID NO:1.
|
Claim 2
A DNA sequence comprising a nucleotide sequence from the 2418th to the 3541st
nucleotide in the nucleotide sequence shown in SEQ ID NO:1.
|
Claim 3
A DNA sequence comprising a nucleotide sequence from the first to the 4120th
nucleotide in the nucleotide sequence shown in SEQ ID NO:2.
|
DNA sequences of a promoter isolated from potato. Partial
and modified sequences of the promoter with cold-inducible promoter activity are
also claimed.
According to the inventors, "cold-inducible" promoter means induced at a
temperature not higher than 6°C and expression of the gene controlled by such
promoter maintained for not less than 5 months.
|
Japan Tobacco
|
|
AU
725727 B2
- Earliest priority - 27 December 1995
- Filed - 26 December 1996
- Granted - 19 October 2000
- Expected expiry - 26 December 1996
|
Title
- Cold-inducible promoter sequences
Claim 1
A DNA sequence having a nucleotide sequence from first to 3546th nucleotide
in the nucleotide sequence shown in SEQ ID. No:1, or a part thereof having a
cold-inducible promoter activity, or a DNA sequence having the same nucleotide
sequence as said DNA sequences except that one or more nucileotides are deleted
or substituted, or one or more nucleotides are inserted or added, which DNA
sequence has a cold-inducible promoter activity.
|
Claim 3
A DNA sequence having a nucleotide sequence from 2418th to 3541st nucleotide
in the nucleotide sequence shown in SEQ ID. No:1, or a part thereof having a
cold-inducible promoter activity, or a DNA sequence having the same nucleotide
sequence as the said DNA sequences except that one or more nucleotides are
deleted or substituted, or one or more nucleotides are inserted or added, which
DNA sequence has a cold-inducible promoter activity.
|
Claim 4
A cold-inducible promoter sequence having a nucleotide sequence from 2418th
to 3541st nucleotide in the nucleotide sequence shown in SEQ ID. No:1, or a DNA
sequence having the same nucleotide sequence as said DNA sequence except that
one or more nucleotides are deleted or substituted, or one or more nucleotides
are inserted or added, which DNA sequence has a cold-inducible promoter
activity.
|
|
Claim 5
A DNA sequence having a nucleotide sequence
from first to 4 120th nucleotide in the nucleotide sequence shown in SEQ ID.
NO:2, or a part thereof having a cold-inducible promoter activity, or a DNA
sequence having the same nucleotide sequence as the said DNA sequences except
that one or more nucleotides are deleted or substituted, or one or more
nucleotides are inserted or added, which DNA sequence has a cold-inducible
promoter activity.
|
|
Claim 7
A probe comprising a DNA fragment having at
least 18 consecutive nucleotides in the region from 45th to 839th
nucleotide in the sequence shown in SEQ ID NO:3 in the Sequence
Listing or a sequence complementary thereto.
|
|
|
EP
812917 B1
- Earliest priority -27 December 1995
- Filed - 26 December 1996
- Granted - 12 May 2005
- Expected expiry - 26 December 2016
|
Title - Cold-inducible promoter sequences
Claim 1
A DNA sequence having a nucleotide sequence from first to 3546th nucleotide
in the nucleotide sequence shown in SEQ ID. No. 1, or a part thereof having a
cold-inducible promoter activity.
|
Claim 2
A DNA sequence having a nucleotide sequence from 2418th to 3541st nucleotide
in the nucleotide sequence shown in SEQ ID. No. 1, or a part thereof having a
cold-inducible promoter activity.
|
Claim 3
A cold-inducible promoter sequence having a nucleotide sequence from 2418th
to 3541st nucleotide in the nucleotide sequence shown in SEQ ID. No. 1.
|
Claim 4
A DNA sequence having a nucleotide sequence from first to 4120th nucleotide
in the nucleotide sequence shown in SEQ ID. No. 2, or a part thereof having a
cold-inducible promoter activity.
|
Claim 5
Use of a DNA fragment having at least 15 consecutive nucleotides in the
region from 45th to 839th nucleotide in the sequence shown in SEQ ID. No. 3 in
the Sequence Listing or a sequence complementary thereto, for the preparation of
a probe for screening cold-inducible promoters.
|
|
|
Remarks
|
The related patents were also granted in Japan (JP
3469902), China (CN 96192162.5) and Canada
(CA 2213991).
|
|
US
6184443
- Earliest priority - 21 October 1994
- Filed - 15 September 1997
- Granted - 6 February 2001
- Expected expiry - 14 September 2017
|
Title - Cold-inducible promoter and tuber-specific promoter
sequence from potato alpha amylase gene
Claim 1
An isolated alpha-amylase promoter having cold-sensitive promoter activity,
having a sequence comprising the 5.5 Kb EcoRI DNA fragment of Solanum
tuberosum from the transformed E. coli strain, DH5alpha-gPAmy 351 (NCIMB
Accession Number 40682).
|
Claim 12
An isolated alpha-amylase promoter having tuber-specific activity and having
a sequence comprising SEQ ID NO: 1.
|
DNA sequences (variant, homologue and fragments) of an isolated
alpha-amylase promoter from potato (Solanum
tuberosum) having cold-sensitive activity. The promoter has tuber tissue
specific activity.
|
Danisco
|
|
Remarks
|
Related patent applications in Europe (EP 787194 A1) and
Canada (CA 2202896 AA) were withdrawn and Lapsed, respectively.
Related Australian (AU 27881/95) application has also lapsed.
|
|
US
5847102
- Earliest priority - 12 April 1995
- Filed - 12 April 1995
- Granted - 8 December 1998
- Expected expiry - 8 December 2015
|
Title - Cold-induced promoter from winter Brassica
napus
Claim 1
An isolated DNA molecule comprising a 5' regulatory region of a low
temperature-responsive gene BN115 from Brassica napus, comprising nucleotides
961-1210 of
SEQ
ID NO:1.
|
Claim 7
An isolated cold inducible promoter, or a functional fragment thereof capable
of regulating the expression of a gene in response to a change in temperature,
said cold inducible promoter comprising nucleotides 1-1271 of
SEQ
ID NO:1.
|
Claim 8
An isolated DNA molecule comprising a sequence of at least 15 contiguous
nucleotides of a 5' regulatory region of a low temperature responsive gene BN115
from Brassica napus, as defined by nucleotides 1-1271 of
SEO
ID NO:1.
|
Claim 13
An isolated enhancer, or a functional fragment thereof each capable of
mediating the expression of a gene under the control of a promoter and said
enhancer or fragment, said enhancer obtained from the regulatory region of BN115
from Brassica napus.
|
Claim 16
An isolated negative regulatory element capable of repressing gene expression
at 22° C., obtained from the regulatory region of BN115 from Brassica napus and
comprising nucleotides 461-623 of
SEQ
ID NO:1.
|
Isolated DNA sequences comprising a promoter and 5' regulatory region
of the low temperature responsive gene BN115 from Brassica
napus. Sequences that hybridize to the promoter sequence and
exhibit low temperature activity are also part of the claimed invention. An
enhancer and a negative regulatory element present in the regulatory region are
also claimed.
|
Canadian Ministry of Agriculture
|
|
CA 2146712
- Earliest priority - 10 April 1995
- Filed - 10 April 1995
- Granted - 25 June 2002
- Expected expiry - 10 April 2015
|
Title - Cold-induced promoter from winter Brassica
napus
The claims are the same as
US
5847102.
|
Note: Patent information was last updated on 12 May 2006.
Search terms
for Japan Tobacco patents: "promoter" in abstract and "Japan Tobacco" in
applicant.
Search terms for Danisco patents: "promoter" in abstract and
"Danisco" in applicant.
Search terms for Canadian Ministry of Agriculture
patents: "promoter" in abstract and "Canadian Ministry" in applicant.
Patent database: PatentLens in combination with INPADOC.
Light-regulated promoters
IP issues
The IP portfolio of Calgene Inc includes a United States
patent that claims the use of light responsive promoters in plant cells. It is
the only patent to date that has been issued with a relatively broad scope. The
individual claims of this patent are discussed in detail below.
The other patents presented in this section relate to light-regulated
promoters isolated from genes of specific organisms. The University of
Warwick in UK, Suntory LTD in Japan and
Mycogen Plant Sciences in the USA have filed patents on the use
of promoters whose expression is induced by light, such as a promoter isolated
from myxobacterium and promoters whose expression is inhibited by light
exposure, such as a promoter isolated from a pea gene.
The following tables contains some bibliographic information about the
patents and patent applications filed by the above mentioned entities and a
brief summary of the most important aspects of the protected invention in the
case of granted patents and of the claims as filed for patent applications.
The patents are classified according to whether the promoters are
Light-inducible or
Light-repressible.
A. Light-inducible promoters
The United States patent
US
5750385 assigned to Calgene Inc, does not
specifically claim a single, individual light-inducible promoter. Rather, the
patent claims the use of a promoter activated by light in order to control
expression of a gene introduced into plants by transformation. In at least some
cases, the phenotype of the tissue expressing the gene is modified.
The specification of the patent discloses promoters such as fruit-specific
and seed specific. Related patents
(US
5420034,
US
5753475 and
US
6281410) have claims directed to these aspects. The claims in the
instant patent are directed only to induced gene expression or modification of
the phenotypic property in transformed plants as a result of light induced
processes in chloroplasts.
The patent specification makes particular reference to the promoter from the
soybean SSU gene (small subunit SSU) of ribulose-1,5-bisphophate-carboxylase,
although the claims are not so limited. Through the use of the soybean SSU
promoter, the expression of the gene under its control can be light-induced.
Thus in the presence of light, its expression is increased, while its expression
is substantially reduced in the absence of light. Moreover, the vector construct
may also include enhancers, operators, activators, or other regions involved
with transcriptional regulation.
The patent claims methods for obtaining a transformed plant with a modified
phenotype or for altering the phenotype of transformed plant tissues containing
chloroplasts using a vector construct comprising:
- a promoter region from a gene that is light-inducible in a chloroplast-co
ntaining tissue,
- DNA sequence of interest, and
- a transcription termination region.
In dependent claims, the promoter is from a soybean SSU gene, and the plant
is soybean, tomato, or rapeseed. Because the claims do not limit the source of
the light-inducible promoter, the Calgene assigned patent has the broadest scope
in the patent landscape surrounding light-inducible promoters.
Calgene Inc. was bought by Monsanto in 1996
and therefore its patents are probably now controlled by
Monsanto
(http://www.monsanto.com/monsanto/layout/about_us/contactus.asp)
Specific Patent Information
|
Patent Number
|
Title, Independent Claims and Summary of Claims
|
Assignee
|
|
US
5750385
- Earliest priority - 17 January 1985
- Filed - 7 June 1995
- Granted 12 May 1998
- Expected expiry - 12 May 2015
|
Title - Methods and compositions for regulated
transcription and expression of heterologous genes.
Claim 1
A method for obtaining a plant having a modified phenotype, said method
comprising:
transforming a host plant cell with a DNA construct under genomic
integration conditions, wherein said construct comprises as
operably linked components in the direction of transcription,
a promoter region obtainable from a gene, wherein transcription of said
gene is light-inducible in a plant chloroplast containing tissue,
a DNA sequence of interest other than the native coding sequence of said
gene, and
a transcription termination region, wherein said components are
functional in a plant cell, whereby said DNA construct becomes integrated into a
genome of said plant cell;
regenerating a plant from said transformed plant cell,
and
growing said plant under conditions whereby said DNA sequence of interest
is expressed and a plant having said modified phenotype is obtained.
|
Claim 2
A method for altering the phenotype of chloroplast containing plant tissue as
distinct from other plant tissue, said method comprising:
growing a plant, wherein said plant comprises cells containing a DNA
construct integrated into their genome, said DNA construct
comprising, in the 5' to 3' direction of transcription,
a transcriptional initiation region from a gene, wherein transcription
of said gene is light-inducible in a plant chloroplast containing tissue,
a DNA sequence of interest other than the coding sequence native to said
transcriptional initiation region, and
a transcriptional termination region, whereby said DNA sequence of
interest is transcribed under transcriptional control of said transcriptional
initiation region and a plant having an altered phenotype is obtained.
|
The claims are generally directed to methods for transforming plants for a
modified phenotype using a DNA construct. The construct comprises a promoter
that provides for light-inducible transcription in either a specific plant
tissue or a plant part. Transcription may be induced during a particular stage
of growth or in response to an external stimulus.
|
Calgene Inc.
|
Note: Patent information was last updated on 11 May 2006. Search terms:
"Promoter" in abstract and "Calgene" in applicant. Patent database: PatentLens
in combination with INPADOC.
University of Warwick had granted patents in Europe
(EP
310619) and Canada (CA 1321563) regarding a light
inducible promoter. The claims are generally related to a DNA sequence
comprising a promoter isolated from myxobacterium. The promoter
is linked to a sequence further containing a restriction site for the insertion
of a desired sequence. A method of producing a polypeptide under the control of
the myxobacterium promoter is also claimed but limited to a transformed
bacterial host.
However, the fact that both the European and the Canadian patents lapsed and
the related Australian patent application (AU 75421/87) also
lapsed suggested that this patent family had been abandoned.
B. Light-repressible promoters
Suntory Ltd.
(http://www.suntory.com/group/suntory_ltd.html)
has patents granted in Australia and New Zealand and pending applications in
Europe and Canada. This patent family mainly claims for a promoter of the
pra2 gene from pea. Mycogen Plant Sciences
(http://www.dowagro.com/mycogen/contact/index.htm)has
two granted United States patents regarding methods for the expression and
enhancing the level of expression of a structural gene in plants by using a
dark- and light-active maize Cab promoter/regulatory system.
Specific Patent Information
|
Patent Number
|
Title, Independent Claims and Summary of Claims
|
Assignee
|
|
AU
765413 B2
- Earliest priority - 12 March 1999
- Filed - 3 March 2000
- granted - 18 September 2003
- Expected expiry - 3 March 2020
|
Title - Photoinhibitory promoter
Claim 1
A DNA fragment containing the sequence of SEQ ID NO: 1 as a
core sequence, whereby expression of a gene placed downstream of said DNA
fragment is repressed in the presence of light.
|
Claim 4
A promoter containing the sequence of SEQ ID NO: 1 as a core
sequence, whereby expression of a gene placed downstream of said promoter is
promoted in the dark but repressed in the presence of light.
|
The claims are drawn to a promoter sequence or a fragment of the promoter
sequence whose ability of controlling the expression of a gene placed downstream
is promoted in the dark but repressed by light.
The promoter was isolated from a small G protein gene
pra2 from pea. The pra2 gene was thought to be
involved in the elongation of stems, at the epicotyl, during germination in the
dark.
|
Suntory Ltd.
|
|
NZ 508103 B
- Earliest priority - 12 March 1999
- Filed - 3 March 2000
- granted - 28 August 2002
- Expected expiry - 3 March 2020
|
Title - Light-repressible promoters
The claims are exactly the same as
AU
765413.
|
|
Remarks
|
Related patent applications are pending in Europe (EP 1077257
A1) and Canada (CA 2328139 A1).
|
|
US
5639952
- Earliest priority - 5 January 1989
- Filed - 7 April 1995
- Granted - 17 June 1997
- Expected expiry - 17 June 2014
|
Title - Dark and light regulated chlorophyll A/B binding
protein promoter-regulatory system
Claim 1
A method for the expression of a structural gene in a plant cell under
conditions of darkness comprising the steps of:
a. transforming said plant cell with a recombinant molecule
comprising a plant, dark- and light-active maize Cab
promoter/regulatory system, wherein said promoter/regulatory system is the
promoter/regulatory system of Cab AB1084 or cross-hybridizes with the
promoter/regulatory system of Cab AB1084 under stringent conditions, and a
heterologous plant-expressible structural gene that is under the regulatory
control of said plant Cab promoter/regulatory system in said plant cell, and
b. maintaining conditions of darkness for expression of said structural gene
in said transformed plant cell.
|
Claim 8
A method for enhancing the level of expression obtained in the dark of a
plant-expressible gene in a plant cell by exposing said plant cell to conditions
of illumination comprising the steps of:
a. transforming said plant cell with a recombinant molecule
comprising a plant, dark- and light-active maize Cab
promoter/regulatory system, wherein said promoter/regulatory system is the
promoter/regulatory system of Cab AB1084 or cross-hybridizes with the
promoter/regulatory system of Cab AB1084 under stringent conditions, and a
heterologous plant-expressible structural gene that is under the regulatory
control of said plant Cab promoter/regulatory system in said plant cell, and
b. maintaining conditions of darkness for expression of said structural gene
in said transformed plant cell, and
c. applying conditions of illumination to said transformed plant cell such
that enhancement by a factor of about 3- to 6-fold in expression of said
structural gene is obtained.
|
Claim 15
A plant grown from a plant cell, wherein said plant cell is transformed with
a recombinant molecule comprising a plant, dark- and light-active maize Cab
promoter/regulatory system, wherein said promoter/regulatory system is the
promoter/regulatory system of Cab AB1084 or cross-hybridizes with the
promoter/regulatory system of Cab AB1084 under stringent conditions, and a
heterologous plant-expressible structural gene that is under the regulatory
control of said plant Cab promoter/regulatory system in said plant cell.
|
This patent mainly claims methods for the expression of a structural gene in
a plant under dark conditions by placing the gene under the control of a dark
and light-active maize chlorophyll a/b (Cab) promoter/regulatory system. The
promoter/regulatory system used is either the Cab AB1084 or cross-hybridizes
under stringent conditions. The expression of the gene under control of the
system is further enhanced by a subsequent period of light. A plant grown from a
plant cell transformed with such promoter/regulatory system is also part of the
claimed invention.
|
Mycogen Plant Sciences
|
|
US
5656496
- Earliest priority - 5 January 1989
- Filed - 5 June 1995
- granted 12 August 1997
- Expected expiry - 12 August 2014
|
Title - Dark and light regulated chlorophyll A/B binding
protein promoter-regulatory system
Claim 1
A recombinant DNA molecule comprising:
(a) a dark- and light-active Cab promoter/regulatory system which
cross-hybridizes with the Cab AB1084 promoter/regulatory system under stringent
conditions, and wherein said promoter/regulatory system
(i) functions to direct the expression of a structural gene under its
control during conditions of darkness and
(ii) is stimulated to direct enhanced expression of said structural gene
under light conditions, and
(b) a heterologous plant-expressible structural gene wherein said structural
gene is placed under the regulatory control of said plant promoter/regulatory
system.
|
This US patent is a division of
US
5639952. It is directed to a dark and light-active
Cab promoter/regulatory system, which cross-hybridizes with the
Cab AB1084. The promoter system operates the same as described
in
US
5639952.
|
Note: Patent information was last updated on 12 May 2006.
Search terms for Suntory patents: "promoter" in title or abstract and "Suntory"
in applicant. Patent database: esp@cenet worldwide.
Search terms for Mycogen patents: "promoter" in abstract and "Mycogen" in
applicant. Patent database: PatentLens in combination with INPADOC.
Chapter 4
Tissue-specific promoters
Summary
As mentioned in the Introduction, there are promoters controlling gene
expression in a tissue-dependent manner and according to the developmental stage
of the plant. The transgenes driven by these type of promoters will only be
expressed in tissues where the transgene product is desired, leaving the rest of
the tissues in the plant unmodified by transgene expression. Tissue-specific
promoters may be induced by endogenous or exogenous factors, so they can be
classified as inducible promoters as well.
Unlike constitutive expression of genes, tissue-specific expression is the
result of several interacting levels of gene regulation. As such, it is then
preferable to use promoters from homologous or closely related plant species to
achieve efficient and reliable expression of transgenes in particular tissues.
This is one of the main reasons for the large amount of tissue-specific
promoters isolated from particular plants and tissues found in both scientific
and patent literature. Patents claiming particular tissue promoters such as
beta-amylase gene or barley hordein gene promoters (for seed gene expression),
tomato pz7 and pz130 gene promoters (for ovary gene expression), tobacco RD2
gene promoter (for root gene expression), banana TRX promoter and melon actin
promoter (for fruit gene expression) and so forth, are the most abundant in the
patent literature.
The number of promoters "tailor-made" (isolated and identified) for the
expression of transgenes in particular tissues and plants is in the hundreds.
Very few patents are drawn to tissue-specific promoters (e.g. seed-specific or a
root-specific promoter) in general terms
The purpose of this section is to present those patents directed to plant
tissue-specific promoters in broad terms. Patents related to particular p
romoters derived from specific genes and plants are beyond the scope of this
paper.
The analysed patents are categorised according to tissue where the promoter
controls the transcription of a gene of interest. These are:
-
Root
promoters: Pioneer Hi-Bred has filed several patent
applications directed to root promoters that enhance or suppress the expression
of a linked gene in root cells. In addition, the invention comprises methods for
the identification and isolation of plant tissue-specific promoters in general.
-
Fruit
promoters: Calgene's large portfolio of patents on
tissue-specific promoters includes fruit specific promoters that control the
expression of genes in mature ovary tissue of a fruit and in the receptacle
tissue of accessory fruits such as strawberry, apple and pear. The genes driven
by the promoters of Calgene's inventions influence fruit development and
ripening.
-
Seed
promoters: Calgene, Sapporo Breweries and the
University of California have granted patents and patent
applications drawn to seed-specific promoters in broad terms. Transcription
cassettes having a seed-specific promoter and recombinant molecules containing a
seed-maturation promoter are part of their inventions.
Root Promoters
Summary
Pioneer Hi-Bred had filed patent applications related to
plant promoters containing elements that drive the expression of genes of
interest in root cells and tissues.
The claims as filed of the patent applications also describe very broad
methods for the isolation and characterization of tissue-specific plant
promoters in general. If the claims as filed are granted without change they
would potentially cover processes for the identification of tissue-specific
promoters regardless of the tissue where the expression of nucleotide sequences
is sought.
Pioneer Hi-Bred has now obviously abandoned this patent, although it is still
pending in Australia.
The Invention
Pioneer Hi-Bred's patent applications in Australia, the U.S.
and Europe cover the following aspects:
- Methods for identifying and isolating tissue-preferred plant promoter
elements in general.
- The elements are not restricted to any particular tissue in the claims as
filed.
- Plant promoters containing root-preferred promoter elements that enhance or
suppress the expression of a linked sequence in root cells.
- Specific sequences of root-promoter elements are spelled out in the claims
as filed. Some plant promoters of the invention have multiple root-preferred
promoter elements.
- Methods for root-preferred expression of genes in plants by transforming a
plant with an expression cassette having a promoter with elements for root
expression; and
- Plant cells stably transformed with DNA construct containing root-preferred
expression elements.
The term "root-preferred" means that the expression driven by a plant
promoter of the invention is selectively enhanced or suppressed in roots in
comparison to other tissues. Root cells and tissues include any part of the
roots, and cover primary, lateral and adventitious roots.
Plants to be transformed with the constructs of the invention are not limited
to any in particular in the claims as filed.
|
Patent No
|
Title, Independent Claims and Summary
|
Applicant
|
|
AU
2001/32896
- Filed - 19 January 2001
- Granted - under examination
- Lapsed on 31 Aug 2006
|
Title - Novel root-preferred promoter elements and method of use
Claim 1
A plant promoter comprising at least one tissue-preferred plant promoter
element, said element identified by:
a) providing a first mixture of
oligonucleotides each comprising a 5' flanking sequence, a central random
sequence, and a 3' flanking sequence;
b) contacting said first mixture with
a second mixture comprising nuclear proteins from a preferred plant tissue under
binding conditions promoting complex formation between said oligonucleotides and
said
proteins;
c) separating said formed complexes
electrophoretically;
d) isolating said separated complexes in ranges of
electrophoretic mobility;
e) amplifying oligonucleotides of said isolated
complexes by polymerase chain reaction utilizing primers to said flanking
sequences ;
f) providing said amplified oligonucleotides from step e) as
the first mixture for a repetition of step a);
g) performing at least a
second cycle of steps b-e with said provided oligonucleotides of step f);
h) assessing for a particular range of electrophoretic mobility and quantity of
complex formation in progressive cycles of step g);
i) isolating
oligonucleotides of a particular range of electrophoretic mobility wherein said
range has increased complex formation in step h);
j) operably linking
individual oligonucleotides of step i) to a promoter that drives expression in a
plant cell, said promoter operably linked to a coding sequence in an expression
cassette;
k) assessing tissue-preferred expression of said coding sequence;
and
I) determining sequence of an oligonucleotide having tissue preferred
expression in step k).
|
|
Claim 5
A plant promoter comprising at least one
root-preferred plant promoter element comprising a nucleotide sequence selected
from the group consisting of:
a) a nucleotide sequence of SEQ ID NO.1, SEQ
ID NO.2, SEQ ID NO.3, SEQ ID NO.4, SEQ ID NO.5, SEQ ID NO.6, SEQ ID NO.7, or SEQ
ID NO.8;
b) a nucleotide sequence that hybridizes under stringent
conditions to a nucleotide sequence of a); and
c) a nucleotide sequence
comprising at least 7 contiguous nucleotides of a sequence of a), wherein said
contiguous nucleotides maintain function of the nucleotide sequence of a).
|
|
Claim 10
A plant promoter comprising at least one
multimeric root-preferred promoter element comprising at least two
root-preferred promoter elements further comprising a nucleotide sequence
selected from the group consisting of :
a) a nucleotide sequence of SEQ ID
NO.1, SEQ ID NO. 2, SEQ ID NO. 3, SEQ ID NO. 4,SEQ ID NO.5, SEQ ID NO. 6,SEQ ID
NO. 7, or SEQ ID NO. 8;
b) a nucleotide sequence that hybridizes under
stringent conditions to a nucleotide sequence of a); and
c) a nucleotide
sequence comprising at least 7 contiguous nucleotides of a sequence of a),
wherein said contiguous nucleotides maintain function of the nucleotide sequence
of a).
|
|
Claim 11
A plant promoter comprising at least one
root-preferred plant promoter element that enhances expression of a coding
sequence operably linked to said promoter, wherein said element comprises a
nucleotide sequence selected from the group consisting of :
a) a nucleotide
sequence of SEQ ID NO.1, SEQ ID NO. 2, SEQ ID NO. 3, SEQ ID NO. 4, SEQ ID NO. 5,
SEQ ID NO. 6,SEQ ID NO. 7, or SEQ ID NO. 8;
b) a nucleotide sequence that
hybridizes under stringent conditions to a nucleotide sequence of a); and
c) a nucleotide sequence comprising at least 7 contiguous nucleotides of a
sequence of a), wherein said contiguous nucleotides maintain function of the
nucleotide sequence of a).
|
Claim 12
A plant promoter comprising at least one root-preferred plant promoter
element that suppresses expression of a coding sequence operably linked to said
promoter, wherein said element comprises a nucleotide sequence selected from the
group consisting of :
a) a nucleotide sequence of SEQ ID NO. 1,SEQ ID NO.
2, SEQ ID NO. 3, SEQ ID NO. 4, SEQ ID NO. 5, SEQ ID NO. 6, SEQ ID NO. 7, or SEQ
ID NO. 8;
b) a nucleotide sequence that hybridizes under stringent
conditions to a nucleotide sequence of a); and
c) a nucleotide sequence
comprising at least 7 contiguous nucleotides of a sequence of a), wherein said
contiguous nucleotides maintain function of the nucleotide sequence of a).
|
|
Claim 13
A transformed plant, or its parts, having
stably incorporated into its genome a DNA construct comprising a plant promoter
operably linked to a coding sequence, said plant promoter comprising at least
one synthetic root-preferred plant promoter element.
|
Claim 19
A transformed plant cell, said plant cell having stably incorporated into its
genome a DNA construct comprising a plant promoter operably linked to a coding
sequence, said plant promoter comprising at least one synthetic root-preferred
plant promoter element.
|
Claim 20
A method for root-preferred expression of a nucleotide coding sequence in a
plant, said method comprising transforming a plant cell with a transformation
vector comprising an expression cassette, said expression cassette comprising a
plant promoter operably linked to said nucleotide coding sequence, said plant
promoter comprising at least one synthetic root-preferred plant promoter
element.
|
Claim 22
A method for identifying and isolating tissue-preferred promoter elements,
said method comprising the steps of :
a) providing a first mixture of
oligonucleotides each comprising a 5' flanking sequence, a central random
sequence, and a 3' flanking sequence;
b) contacting said first mixture with
a second mixture comprising nuclear proteins from a preferred plant tissue under
binding conditions promoting complex formation between said oligonucleotides and
said proteins;
c) separating said formed complexes
electrophoretically;
d) isolating said separated complexes in ranges of
electrophoretic mobility;
e) amplifying oligonucleotides of said isolated
complexes by polymerase chain reaction utilizing primers to said flanking
sequences;
f) providing said amplified oligonucleotides from step e) as the
first mixture for a repetition of step a);
g) performing at least a second
cycle of steps b-e with said provided oligonucleotides of step f) ;
h)
assessing for a particular range of electrophoretic mobility and quantity of
complex formation in progressive cycles of step g);
i) isolating by
cloning, individual oligonucleotides of a particular range of electrophoretic
mobility wherein said range has increased complex formation in step h);
j)
simultaneous with step i) or as an individual step, operably linking isolated
individual oligonucleotides of step i) to a promoter that drives expression in a
plant cell, said promoter operably linked to a coding sequence in an expression
cassette;
k) assessing tissue-preferred expression of said coding sequence;
and
l) determining sequence of an oligonucleotide having tissue preferred
expression in step k).
|
|
Pioneer Hi-Bred
|
|
Remarks
|
Related application in the United States
(US
2001/047525 A1) has been expressly abandoned. Applications
in Europe (EP 1248850 A2) and Canada (CA
2390819) are also lapsed.
|
|
Fruit Promoters
Among the large patent portfolio of Calgene on
tissue-specific promoters, there are three main patent families containing
granted patents directed to fruit-specific regulatory regions. Patents and
patent applications that were assigned to Calgene may now be held by Monsanto,
to which any inquiries about licensing should be directed
(http://www.monsanto.com/monsanto/layout/about_us/contactus.asp).
Patents of one of the patent families are drawn to DNA constructs containing
promoters "preferentially expressed in fruit tissues". Although the basic
patents in the patent family are involved in the expression of genes regulating
fruit ripening and drive the expression of genes of interest in mature ovaries,
the definitions are broad, and examples in which seed-specific expression is
cited suggest that tissues such as seed, fruit integument, cotton fibers
and so on would be construed as "fruit tissues". In addition, general
methods to regulate the fruit phenotype are part of the patented inventions.
In the other two patent families, the definitions are more specific. One
relates primarily to cotton fiber production, although it contains some broader
claims. In the second, the promoters of the invention are expressed in
receptacle tissue, a flower part that makes most of the fleshy
tissue in accessory fruits such as strawberry, apple and pear. The genes driven
by the promoter influence fruit development, maturation and ripening. Some
analysis of this patent family is provided below.
Broadest patent family
The claims of the United States patent are drawn to methods
for altering the phenotype of the fruit tissue of a plant transformed with DNA
constructs comprising:
- a promoter preferentially expressed in a fruit tissue (note that this does
not mean "fruit-specific");
- a DNA sequence of interest different from the native gene of the promoter
(note that this need not be a "gene"--could be antisense or RNAi); and
- a transcriptional termination region.
Unlike the European patent, the genes from which the fruit-specific promoters
are obtained are not limited to anthesis process or fruit maturation and
ripening. The promoters are from genes preferentially transcribed in fruit
tissue. Because there is no limitation in the fruit gene promoter used in the
DNA construct to alter the phenotype of a fruit, the invention claimed in the
United States patent, although directed to methods, is broader in scope than in
the Canadian and European patents. Also, the claimed methods are described in
very broad terms.
Calgene's European patent claims:
- A DNA construct comprising a fruit transcriptional initiation region from a
gene that becomes active during anthesis (period during which a flower is fully
open and functional), remains active until the ripe period and is transcribed in
mature ovary tissue. The construct includes a gene of interest under the control
of such transcriptional region and a transcriptional termination region.
- A DNA construct having a fruit promoter of a plant storage protein that
becomes and remains active during the stages described above.
- A method for the modification of the phenotype of a tomato fruit. The sort
of phenotypic modification is not specified in the claim.
- A tomato cell is transformed with a DNA construct as described above. A
tomato plant is regenerated and grown from the transformed cell.
|
Patent Number
|
Title, Independent Claims and
Summary of Claims
|
Assignee
|
|
US
4943674
- Earliest priority -17 January 1985
- Filed - 26 May 1987
- Granted - 24 July 1990
- Expected expiry - 24 July 2007
|
Title - Fruit specific transcriptional factors
Claim 1
A DNA construct comprising in the direction of
transcription,
a tomato 2All transcriptional initiation region joined
to a DNA sequence of interest, wherein said DNA sequence of interest is other
than the wild-type sequence and is under the transcriptional regulation of said
2All initiation region and
a transcriptional termination region.
|
Claim 5
A DNA construct comprising in the direction of
transcription,
a tomato 2All transcriptional initiation region joined
to a DNA sequence of interest, wherein said DNA sequence is other than the
wild-type sequence and comprises a unique restriction site for
insertion of a second DNA sequence of interest to be under the transcriptional
regulation of said 2All initiation region, and
a transcriptional
termination region.
|
Claim 9
A method for modifying the phenotype of tomato fruit said method
comprising:
transforming a tomato plant cell with a
DNA construct under genomic integration conditions, wherein said DNA construct
comprises in the direction of transcription,
a
tomato 2All transcriptional initiation region jointed to a DNA sequence other
than the wild-type sequence and capable of modifying the phenotype of fruit
cells upon transcription, wherein said DNA sequence is under the transcriptional
regulation of said initiation region and
a transcriptional
termination region, whereby said DNA construct becomes integrated into the
genome of said tomato plant cell; regenerating a plant from said
transformed tomato plant cell; and
growing said plant to produce
tomato fruit of the modified phenotype.
|
Claim 12
Tomato fruit comprising a DNA construct
comprising in the direction of transcription,
a
tomato 2All transcriptional initiation region joined to a DNA sequence of
interest, wherein said DNA sequence is other than the wild-type sequence and is
under the transcriptional regulation of said 2All initiation region, and
a transcriptional termination region.
|
Claim 13
A tomato plant comprising: a DNA construct
comprising in the direction of transcription,
a
tomato 2All transcriptional initiation region joined to a DNA sequence of
interest, wherein said DNA sequence is other than the wild-type sequence and is
under the transcriptional regulation of said 2All initiation region, and
a transciptional termination region.
|
Claim 14
A tomato plant comprising: a DNA construct
comprising in the direction of transcription,
a
tomato 2All transcriptional initiation region joined to a DNA sequence of
interest, wherein said DNA sequence is other than the wild-type sequence and
comprises a unique restriction site for insertion of a second
DNA sequence of interest to be under the transcriptional regulation of said 2All
initiation region, and
a transciptional termination region.
|
|
Calgene
Inc.
|
|
US
5753475
- Earliest priority -17 January 1985
- Filed - 10 August 1993
- Granted - 19 May 1998
- Expected expiry - 19 May 2015
|
Title - Methods and compositions for regulated transcription
and expression of heterologous genes
Claim 1
A method for obtaining a plant having a regulatable phenotype, said method
comprising:
transforming a host plant cell with a DNA
construct under genomic integration conditions, wherein said construct
comprises as operably linked components in the direction of
transcription,
a promoter region obtainable from a gene, wherein
transcription of said gene is preferentially regulated in a plant fruit
tissue,
a DNA sequence of interest other than the native coding
sequence of said gene, and
a transcription termination region,
wherein said components are functional in a plant cell, whereby said DNA
construct becomes integrated into a genome of said plant cell;
regenerating a plant from said transformed plant cell, and
growing
said plant under conditions whereby said DNA sequence of interest is expressed
and a plant having said regulatable phenotype is obtained.
|
Claim 2
A method for altering the phenotype of fruit tissue as distinct from other
plant tissue, said method comprising:
growing a
plant, wherein said plant comprises cells containing a DNA
construct integrated into their genome, said DNA construct
comprising, in the 5' to 3' direction of transcription,
a transcriptional initiation region from a gene, wherein transcription
of said gene is preferentially regulated in a plant fruit tissue,
a DNA sequence of interest other than the coding sequence native to said
transcriptional initiation region, and
a transcriptional
termination region, whereby said DNA sequence of interest is transcribed under
transcriptional control of said transcriptional initiation region and a plant
having an altered phenotype is obtained.
|
|
Claim 5
A method for modifying the genotype of a plant
to impart a desired characteristic to fruit as distinct from other plant tissue,
said method comprising:
transforming under genomic
integration conditions, a host plant cell with a DNA construct
comprising in the 5' to 3' direction of transcription,
a transcriptional initiation region from a gene, wherein transcription
of said gene is preferentially regulated in a plant fruit tissue,
a DNA sequence of interest other than the native coding sequence of said gene,
and
a transcriptional termination region, whereby said DNA
construct becomes integrated into the genome of said plant cell;
regenerating a plant from said transformed host cell; and
growing said
plant to produce fruit having a modified genotype.
|
Claim 9
A method for modifying transcription in fruit tissue as distinct from other
plant tissue, said method comprising:
growing a plant
capable of developing fruit tissue under conditions to produce fruit, wherein
said plant comprises cells containing a DNA construct
integrated into their genome, said DNA construct comprising, in
the 5' to 3' direction of transcription,
a fruit-specific
transcriptional initiation region,
a DNA sequence of interest
other than the coding sequence native to said transcriptional initiation region,
and
a transcriptional termination region, whereby said DNA
sequence of interest is transcribed under transcriptional control of said
fruit-specific transcription initiation region.
|
|
Claim 12
A method to selectively express a heterologous
DNA sequence of interest in fruit tissue as distinct from other plant tissue,
said method comprising:
growing a plant capable of developing fruit tissue under conditions to
produce fruit, wherein said plant comprises cells having a genomically
integrated DNA construct comprising, as operably linked
components in the 5' to 3' direction of transcription,
a
fruit-specific transcriptional initiation region and a translational initiation
region,
a DNA sequence of interest other than the coding sequence
native to said transcriptional initiation region,
a
transcriptional termination region downstream of said DNA sequence of interest,
whereby said DNA sequence of interest is expressed under control of said
fruit-specific transcriptional and translational initiation region.
|
This patent is also a Continuation in part of
US
5420034 (see the
Seed promoter
section).
|
|
EP
316441 B1
- Earliest priority - 26 May 1987
- Filed - 26 May 1988
- Granted - 5 December 2001
- Expected expiry - 26 May 2008
|
Title - Fruit-specific transcriptional factors
|
Claim 1
A DNA construct comprising in
the direction of transcription,
a fruit-specific transcriptional
initiation region from a gene which becomes active at or immediately after
anthesis and remains active at least until the ripe period, and which is
transcribed in mature ovary tissue, joined to a DNA sequence of interest other
than the wild-type sequence associated with said initiation region, wherein said
DNA sequence of interest is under the transcriptional regulation of said
initiation region and
a transcriptional termination region.
|
|
Claim 7
A DNA construct comprising in
the direction of transcription,
a fruit-specific transcriptional
initiation region of a plant storage protein which becomes active at or
immediately after anthesis and remains active at least until the ripe period and
which is transcribed in mature ovary tissue, joined to a DNA sequence other than
the wild-type sequence, wherein said sequence comprises a
unique restriction site for insertion of a sequence of interest to be under the
transcriptional regulation of said initiation region, and
a
transcriptional termination region.
|
|
Claim 12
A method for specifically modifying the
phenotype of fruit substantially distinct from other plant tissue, said method
comprising:
transforming a plant cell with a DNA
construct under genomic integration conditions, wherein said DNA construct
comprises in the direction of transcription,
a
fruit-specific transcriptional initiation region which becomes active at or
immediately after anthesis, and remains active at least until the ripe period
and which is transcribed preferentially in mature ovary tissue, joined to a DNA
sequence other than the wild-type sequence and capable of modfying the phenotype
of fruit cells upon transcription, wherein said sequence is under the
transcriptional regulation of said initiation region, and
a
transcriptional termination region, whereby said DNA construct becomes
integrated into the genome of said plant cell;
regenerating a plant
from said transformed plant cell; and
growing said plant to produce
fruit of the modified phenotype.
|
|
|
Remarks
|
A related Canadian application (CA 1338827) has lapsed and a
related patent was granted in New Zealand (NZ 224787). There
is also a related patent in Australia. Related patent applications also filed in
Israel (IL 86515 A0), and Japan (JP 2500163
T2). Related patent application in China (CN 1036305
A) was withdrawn 20 March 1991.
|
Note: Patent information was last updated on 15 May 2006. Search terms:
"promoter" in abstract and "Calgene" in applicant. Patent database: PatentLens
in combination with INPADOC.
Cotton fiber patent family
As mentioned above, Cotton (Gossypium hirsutum) fiber can be
construed as a type of "fruit tissue" in broader botanical terms. Calgene has a
United States patent directed to a promoter from cotton expansin gene. The
cotton expansin gene is expressed in developing fiber and, according to the
specification, the promoter of the cotton expansin gene can be used to drive a
gene of interest in developing cotton fiber for modifying cotton
fiber phenotypes.
|
Patent Number
|
Title, Independent Claims and
Summary of Claims
|
Assignee
|
|
US
6566586
- Earliest priority - 7 January 1997
- Filed - 7 January 1998
- Granted - 20 May 2003
- Expected expiry - 7 January 2018
|
Title - Cotton expansin promoter sequence
Claim 1
An isolated DNA sequence comprising the sequence of
SEQ
ID NO: 1.
|
The only independent claim is drawn to an isolated DNA sequence comprising
the 2614 bp cotton expansin promoter sequence. A recombinant DNA construct
comprising the promoter sequence and a plant comprising a plant cell comprising
the DNA construct are also claimed in the dependent claims.
|
Calgene LLC
|
|
Remarks
|
Related European application (EP 968292 A1) was withdraw and
the application in Australia (AU 57322/98) also lapsed.
|
Note: Patent information was last updated on 18 May 2006. Search terms:
"promoter" in abstract and "Calgene" in applicant. Patent database: PatentLens
in combination with INPADOC.
Receptacle patent family
A European patent application and a granted United States patent filed by
Calgene are directed to fruit promoters in general driving the expression of
genes in the receptacle tissue of a fruit.
Specific Patent Information
|
Patent Number
|
Title, Summary of Claims and Independent Claims
|
Assignee
|
|
EP
973922
- Earliest priority- 6 February 1998
- Filed - 4 February 1999
- Withdrawn - 7 January 2004
|
Title - Strawberry fruit promoters for gene expression
The claims as filed of the European patent application are broader in this
case, and recite:
- DNA constructs comprising a transcriptional factor driving the expression of
a heterologous gene in the receptacle tissue of a fruit. The expression of the
gene of interest is either:
- increased during fruit ripening or
- decreased during fruit development and maturation.
|
Calgene Inc.
|
|
US
6043410
- Earliest priority- 6 February 1998
- Filed - 6 February 1998
- Granted - 28 March 2000
- Expected expiry - 6 February 2018
|
Title - Strawberry fruit promoters for gene expression
Claim 1
A DNA construct comprising the promoter sequence from RJ39, isolated from
Fragaria, operably-linked to a heterologous DNA coding sequence of interest.
|
|
|
CA
2285465
- Filed - 4 February 1999
- Expiration - Dead application 4 February 2004
|
Title - Strawberry fruit promoters for gene expression
Dead application
|
Note: Patent information was last updated on 15 May 2006. Search terms:
"promoter" in abstract and "Calgene" in applicant. Patent database: PatentLens
in combination with INPADOC.
Seed Promoters
PLEASE NOTE that patent claims drawn to promoters that are active
constitutively in plants, promoters that are active in "fruits" or in
"reproductive parts" may be construed to cover seed-preferred promoters.
Patent claims to particular sequences or to promoters that drive
particular seed-specific genes in certain species (such as phaseolin or napin)
may also be applicable, even though they may not mention the word "seed".
Thus, the list below mentioning a few patents claiming promoters that are
broadly seed-specific is NOT a comprehensive list of promoters that are covered
by patent claims.
Calgene, Sapporo Breweries and the
University of California
have filings drawn to seed-specific promoters in broad terms, listed below.
A patent family with several patents granted to Calgene, directed to a
transcription cassette having a seed-specific promoter, is noted below, and see
also the fruit-specific claims of Calgene elsewhere in this landscape, which may
be construed to apply to seeds.
Patents and patent applications that were assigned to Calgene may now be held
by Monsanto, to which any inquiries about licensing should be directed.
The granted United States patents
US
5420034 and
US
5608152 are directed to promoters isolated from specific
seed genes (i.e. napin gene) and plants (i.e. Brassica). There are
three other granted patents in this patent family, and patent applications in
the same patent family may still be pending.
Note that patent claims are not granted the same way in every country, and
this patent family presents a perfect example of that. The Australian granted
patent claims are broad:
- A seed comprising a transcription cassette containing:
- a seed-specific transcriptional initiation region;
- a sequence of interest other than the native sequence regulated by the
transcriptional region; and
- a transcriptional termination region.
- A transcription construct comprising a polylinker with at least two
restriction sites for the insertion of DNA sequences of interest under the
control of a seed-specific promoter.
- A method to modify the genotype of a seed by the use of a transcription
cassette as described above.
|
Patent Number
|
Title, Independent Claims and Summary
|
Assignee
|
|
EP
255378 B2
- Earliest priority - 31 July 1986
- Filed - 30 July 1987
- Granted - 13 April 1994
- Expected expiry - 30 July 2007
|
Title - Seed-specific transcriptional regulation
Claim 1
A DNA construct comprising in the 5' to 3' direction of
transcription:
a napin transcriptional initiation region, joined to a
DNA sequence of interest other than
(i) a DNA sequence encoding
napin or
(ii) a DNA sequence encoding a mammalian protein or
peptide or mammalian viral pathogen peptide or protein; and
a
transcription termination region.
|
|
Claim 6
A method comprising the production of a DNA
construct comprising in the 5' to 3' direction of
transcription:
a napin transcriptional initiation region wherein said
napin transcriptional initiation region is free from the native DNA sequence
under the regulatory control of said initiation region, joined to a cloning
site, and
a transcriptional termination region; provided that said construct does
not comprise a DNA sequence encoding a mammalian peptide or protein or mammalian
viral pathogen peptide or protein operably linked to said napin transcriptional
initiation region.
|
|
Claim 8
A method of modifying the genotype of a plant
to impart a desired characteristic to seed as distinct from other plant said
method comprising:
transforming a host plant cell
under genomic integration conditions with a DNA construct
comprising in the 5' to 3' direction of transcription:
a seed specific napin transcriptional initiation region, joined to a DNA
sequence of interest other than (a) a DNA sequence encoding a napin or (b) a DNA
sequence encoding a mammalian peptide or protein or a mammalian viral pathogen
peptide or protein; and
a transcriptional termination region; and
growing said plant to produce seed.
|
|
Claim 9
A method for specifically modifying the
phenotype of seed as distinct from other plant tissue, said method
comprising:
(i) transforming a host plant cell under genomic
integration conditions with a DNA construct comprising in the 5' to 3' direction
of transcription:
a seed specific napin transcriptional initiation
region, joined to a DNA sequence of interest other than (a) a DNA sequence
encoding napin or (b) a DNA sequence encoding a mammalian peptide or protein or
a mammalian viral pathogen peptide or protein; and
a
transcriptional termination region; and
(ii) growing a plant under
conditions to produce seed, said plant being comprised of cells capable of
developingseed tissue and said cells having integrated in their genome said DNA
construct.
|
|
Claim 10
A method comprising the production of a DNA
construct comprising in the 5' to 3' direction of transcription:
a seed
specific transcriptional initiation region which is from other than the bean
phaseolin promoter;
a DNA sequence other than the natural coding
sequence joined to said initiation region, wherein said sequence encodes an acyl
carrier protein; and
a transcriptional termination region.
|
|
Claim12
A method for modifying the genotype of a plant
to impart a desired characteristic to seed as distinct from other plant tissues
said method comprising:
transforming a host plant cell under genomic
integration conditions with a DNA construct comprising in the 5' to 3' direction
of transcription:
a seed specific transcriptional initiation
region;
a DNA sequence encoding an acyl carrier protein joined to
said initiation region; and
a transcriptional termination region;
and
growing said plant to produce seed.
|
|
Claim 13
A method for specifically modifying the
phenotype of seed as distinct from other plant tissue, said method comprising:
(i) transforming a host plant cell under genomic integration
conditions with a DNA construct comprising in the 5' to 3' direction of
transcription:
a seed specific transcriptional initiation
region;
a DNA sequence encoding an acyl carrier protein joined to
said initiation region; and
a transcriptional termination region;
and
(ii) growing a plant under conditions to produce seed, said plant
being comprised of cells capable of developing seed tissue and said cells having
integrated in their genome said DNA construct.
|
|
Calgene Inc.
|
|
US
5420034
- Earliest priority - 31 July 1986
- Filed - 8 August 1991
- Granted - 30 May 1995
- Expected expiry - 30 May 2012
|
Title - Seed-specific transcriptional regulation
|
Claim 1
A DNA construct comprising: in
the 5' to 3' direction of transcription,
a transcriptional initiation region from a gene which encodes a product
preferentially expressed in a plant seed cell as compared with other plant
cells,
a DNA sequence of interest other than the native coding
sequence of said gene, and
a transcriptional termination region,
wherein said gene is a napin gene, an acyl carrier protein gene or an EA9 gene.
|
|
Claim 8
An expression cassette
comprising: in the 5'-3' direction of transcription,
a seed-specific transcriptional initiation region wherein said
transcriptional initiation region is free from the native DNA sequence under the
regulatory control of said initiation region,
a cloning site, and
a transcriptional termination region, wherein said transcriptional
initiation region is from a napin gene, an acyl carrier protein gene or an EA9
gene.
|
|
Claim 9
An expression cassette
comprising: in the 5'-3' direction of transcription,
a transcriptional initiation region and ribosome binding site from a gene
expressed in a seed embryo or a seed coat cell or from a gene encoding a seed
storage protein,
a linker or polylinker having one or a plurality of
restriction sites for insertion of a gene to be expressed under transcriptional
control of said transcriptional initiation region, and
a
transcriptional termination region, wherein said transcriptional initiation
region and said ribosome binding site are from a napin gene, an acyl carrier
protein gene or an EA9 gene.
|
|
|
US
5608152
- Earliest priority - 31 July 1986
- Filed - 30 May 1995
- Granted - 4 March 1997
- Expected expiry - 4 March 2014
|
Title - Seed-specific transcriptional regulation
Claim 1
A Brassica plant comprising: a DNA construct
comprising, in the 5' to 3' direction of transcription,
a transcriptional initiation region from a gene that encodes a product
preferentially expressed in a plant seed cell as compared to other plant cells,
a DNA sequence of interest other than the native coding sequence of
said gene, and
a transcriptional termination region, wherein said gene
is a napin gene, an acyl carrier protein gene or an EA9 gene.
|
Claim 8
A Brassica seed comprising: a DNA construct
comprising, in the 5' to 3' direction of transcription,
a transcriptional initiation region from a gene that encodes a product
preferentially expressed in a plant seed cell as compared to other plant cells,
a DNA sequence of interest other than the native coding sequence of
said gene, and
a transcriptional termination region, wherein said gene
is a napin gene, an acyl carrier protein gene or an EA9 gene.
|
This patent is a Division of
US
5420034.
|
|
Remarks
|
Related patents granted in Australia (AU 612326 B2) and New
Zealand (NZ 221259). Related application filed in China
(CN 87106120) has been withdrawn.
|
Note: Patent information was last updated on 15 May 2006. Search terms:
"seed" in abstract and "Calgene" in applicant. Patent database: PatentLens in
combination with INPADOC.
Sapporo Breweries' patents and applications
The granted US and Australian patents claim an isolated barley β-amylase
promoter sequence. However, the independent claims as filed in the European and
Canadian patent applications are very broad as they recite a promoter capable of
expressing an introduced gene in plant seeds. There is no limitation in the gene
source of the seed promoter. In dependent claims the promoter is derived from a
beta-amylase gene from barley. It remains to be seen whether the independent
claims will be granted as filed.
|
Patent Number
|
Title, Independent Claims and Summary
|
Assignee
|
|
US
5952489
- Earliest priority - 5 May 1995
- Filed - 4 March 1997
- Granted - 14 September 1999
- Expected expiry - 4 March 2017
|
Title - Tissue-specific promoter
|
Claim 1
An isolated barley β-amylase promoter
comprising
SEQ
ID NO: 1.
|
|
Sapporo Breweries Ltd.
|
|
AU
717055 B2
- Earliest priority - 5 May 1995
- Filed - 4 March 1997
- Granted - 14 September 1999
- Expected expiry - 4 March 2017
|
Title - Tissue-specific promoter
|
Claim 1
An isolated barley β-amylase promoter
comprising a nucleic acid
sequence of
SEQ
ID NO: 1, or a nucleic acid sequence of
SEQ
ID NO: 1 in which one or more bases are deleted, substituted or added to
said sequence and which has promoter activity in plant seeds.
|
|
|
EP 781849 A1
- Earliest priority - 5 May 1995
- Filed - 5 July 1996
- Granted - Pending
- Expected expiry - N/A
|
Title - Tissue-specific promoter
|
Claim 1
A promoter capable of expressing an introduced gene in plant
seeds
|
|
|
Remarks
|
The related patent application in Canada (CA 2199158) is
also pending.
|
Note: Patent information was last updated on 15 May 2006. Search terms:
"seed" in abstract and "Sapporo" in applicant. Patent database: PatentLens and
esp@cenet in combination with INPADOC.
University of
California's patents and applications
Only independent claim 1 as filed of the European patent application is
relevant for the present paper as it describes in general terms a recombinant
nucleic acid molecule having a seed-maturation specific promoter. The promoter
drives the expression of a protein in a subcellular compartment due to the
presence of a signal peptide that targets the polypeptide to an intracellular
body.
The other independent claims are more specific and describe promoter and
signal sequences derived from barley hordein storage protein. It is important
to note that claims from pending applications may still be granted.
|
Patent Number
|
Title, Independent Claims and Summary
|
Assignee
|
|
US
6642437
- Earliest priority - 30 September 1997
- Filed - 30 September 1998
- Granted - 4 November 2003
- Expected expiry - 30 September 2018
|
Title - Production of proteins in plant seeds
|
Claim 1
A method for producing seeds containing a
selected heterologous protein which is not a seed-storage protein,
comprising the steps of:
(a) stably transforming monocot plant cells with a chimeric gene
having:
(i) a transcriptional regulatory region
from the gene of a maturation specific monocot storage protein selected from the
group consisting of rice glutelins, rice oryzins, rice prolamines, barley
hordeins, oat glutelins, and sorghum kafirins, millet pennisetins, and rye
secalins,
(ii) operably linked to said transcriptional regulatory
region, a first DNA sequence encoding a monocot seed-specific N-terminal leader
sequence capable of targeting a linked polypeptide to a protein storage body in
monocot seeds, and
(iii) a second DNA sequence encoding such
selected non-seed-storage heterologous protein, and linked in translation frame
with the first sequence, such that the first and second sequences encode a
fusion protein composed of the selected heterologous non-seed-storage protein
and an N-terminal leader sequence,
(b) cultivating plants containing
the transformed plant cells under seed-maturation conditions, wherein the
expression of the non-seed storage heterologous protein is at least twice that
observed with an equivalent chimeric gene lacking the second DNA sequence
encoding a monocot seed-specific N-terminal leader sequence, and
(c)
harvesting seeds from the cultivated plants.
|
|
The Regents of the University of
California
|
|
AU
746032 B2
- Earliest priority - 30 September 1997
- Filed - 30 September 1998
- Granted - 11 April 2002
- Expected expiry - 30 September 2017
|
Title - Production of proteins in plant seeds
|
Claim 1
A method of producing a selected heterologous
protein which is not a seed-storage protein, comprising the
steps of:
(a) stably transforming a monocot plant cell with a
chimeric gene comprising:
(i) a transcriptional
regulatory region from the gene of a maturation specific monocot storage protein
selected from the group consisting of rice glutelins, oryzins, and prolamines,
barley hordeins, wheat gliadins and glutenins, maize zeins and glutelines, oat
glutelins, and sorghum kafirins, millet pennisetins, and rye secaliiis,
(ii) operably linked to said transcriptional regulatory region, a first
DNA sequence encoding a monocot seed-specific leader sequence capable of
targeting a linked polypeptide to a protein storage body in monocot seeds, and
(iii) a second DNA sequence encoding the selected heterologous
protein, and linked in translation frame with the first sequence, such that the
first and second sequences encode a fusion protein composed of the selected
protein and an N-terminal leader sequence, and
(b) cultivating plants
containing the cell under seed-maturation conditions to produce the selected
heterologous protein.
|
|
Claim 5
A method of producing a transformed plant
comprising the steps of:
a) stably transforming a
monocot plant cell with a chimeric gene comprising:
(i) a transcriptional regulatory region from the gene of a maturation
specific monocot storage protein selected from the group consisting of rice
glutelins, oryzins, and prolamines, barley hordeins, wheat gliadins and
glutenins, maize zeins and glutelines, oat glutelins, and sorghum kafirins,
millet pennisetins, and rye secalins,
(ii) operably linked to said
transcriptional regulatory region, a first DNA sequence encoding a monocot
seed-specific leader sequence capable of targeting a linked polypeptide to a
protein storage body in monocot seeds, and
(iii) a second DNA
sequence encoding a selected heterologous protein which is not a seed-storage
protein, and linked in translation frame with the first sequence, such that the
first and second sequences encode a fusion protein composed of the selected
protein and an N-terminal leader sequence; and
(b) cultivating a plant
containing the cell.
|
|
|
EP 1019517 A2
- Earliest priority - 30 September 1997
- Filed - 30 September 1998
- Granted - Pending
- Expected expiry - N/A
|
Title - Production of proteins in plant seeds
|
Claim 1
A recombinant nucleic acid molecule having a
structure P-X or P-SS-X, wherein X is a nucleic acid molecule encoding a
polypeptide, P is a seed maturation-specific promoter, and SS is a signal
sequence that targets a linked polypeptide to an intracellular body.
|
|
Claim 12
A recombinant nucleic acid molecule having a
structure Ph-hSS-X, wherein Ph is a hordein promoter, hSS is a hordein signal
sequence, and X is a nucleic acid molecule encoding a polypeptide, and where Ph,
hSS and X are operably inked.
|
|
Claim 30
A method of producing a stably transformed
monocotyledenous plant expressing a selected polypeptide in seeds of the plant,
comprising:
(a) placing an immature zygotic embryo of
the plant on plant growth medium comprising maltose as a sugar
source an auxin at a concentration of about 0.1 mg/L to about 5 mg/L, a
cytokinin at a concentration of 0 mg/L to about 5 mg/L and copper at a
concentration of about 0.1uM to about 50uM, and incubating in dim light
conditions so as to form green regenerative tissue;
(b) introducing a
nucleic acid molecule into the tissue to produce transformed tissue, wherein the
nucleic acid molecule has a structure Ph-hSS-X, wherein Ph is a hordein
promoter, hSS is a hordein signal sequence, and X is a nucleic acid molecule
encoding the selected polypeptide, and where Ph, hSS and X are operably
linked;
(c) incubating the transformed tissue on the plant growth
medium such that green structures are observed on the transformed
material;
(d) regenerating at least one transformed plant from the
green structures; and
(f) growing the transformed plant to produce
seed.
|
|
Claim 35
A method of producing a stably transformed
monocotyledenous plant expressing a selected polypeptide in seeds of the plant,
comprising:
(a) placing an immature zygotic embryo of
the plant on plant growth medium comprising maltose as a sugar
source, an auxin at a concentration of about 0.1 mg/L to about 5 mg/L, a
cytokinin at a concentration of0 mg/L to about 5 mg/L and copper at a
concentration of about0.1 uM to about 50ltM, and incubating in dim light
conditions so as to form green regenerative tissue;
(b) introducing a
nucleic acid molecule into the tissue by to produce transformed tissue, wherein
the nucleic acid molecule has a structure Ph-X, wherein Ph is a hordein promoter
and X is a nucleic acid molecule encoding the selected polypeptide, and where Ph
and X are operably linked;
(c) incubating the transformed tissue on the
plant growth medium such that green structures are observed on the transformed
material;
(d) regenerating at least one transformed plant from the
green structures; and
(f) growing the transformed plant to produce
seed.
|
|
Claim 39
A method of producing a stably transformed
monocotyledenous plant expressing a selected polypeptide in seeds of the plant,
comprising:
(a) placing an immature zygotic embryo of
the plant on plant growth medium comprising maltose as a sugar
source, an auxin at a concentration of about 0.1 mg/L to about 5 mg/L, a
cytokinin at a concentration of0 mg/L to about 5 mg/L and copper at a
concentration of about 0. 11M to about 50I1M, and incubating in dim light
conditions so as to form green regenerative tissue;
(b) introducing a
nucleic acid molecule into the tissue by to produce transformed tissue, wherein
the nucleic acid molecule has a structure P - X or P - SS - wherein X is a
nucleic acid molecule encoding a polypeptide, P is a seed maturation-specific
promoter, and SS is a signal sequence that targets a linked polypeptide to an
intracellular body;
(c) incubating the transformed tissue on the plant
growth medium such that green structures are observed on the transformed
material;
(d) regenerating at least one transformed plant from the
green structures; and
(f) growing the transformed plant to produce
seed.
|
|
|
Remarks
|
An application as a Continuation of
US
6642437, which has exactly the same claims as (EP
1019517), is pending in the United States
(US
2004/88754 A1). Related applications are also pending in Canada
(CA 2305628) and Japan (JP 2001518305)
|
Note: Patent information was last updated on 15 May 2006. Search terms:
"seed" in abstract and "University of California" in applicant. Patent database:
PatentLens and esp@cenet in combination with INPADOC.
Appendix 1
Claims
Opine Promoters
The CaMv promoters: 35S and 19S
Plant Ubiquitin Promoter
System
Maize alcohol dehydrogenase 1 (Adh-1)
promoter
Synthetic promoters
1. Promoters and UAS from opine synthase genes
Patents granted to Biotechnology Research And Development Corporation &
Purdue Research Foundation
Actual granted independent claims
|
US
5955646
|
|
Claim 1
A cassette for inducible expression of a
foreign gene comprising said foreign gene operably linked to a regulatory region
comprising:
a) a promoter derived from a mannopine synthase gene of Agrobacterium
tumefaciens,
b) an upstream activating sequence derived from a
mannopine synthase gene of Agrobacterium tumefaciens, and
c) at
least one upstream activating sequence derived from an octopine synthase gene of
Agrobacterium tumefaciens.
|
|
Claim 2
A method for nematode inducible expression of
a foreign gene in a plant, comprising:
a) linking said foreign gene to a regulatory region comprising a promoter
derived from a mannopine synthase gene of Agrobacterium tumefaciens
comprising 138 bases upstream of the transcription initiation site, and an
upstream activating sequence derived from a mannopine synthase gene of
Agrobacterium tumefaciens;
b) inserting said foreign gene and said
regulatory region in said plant, wherein expression is induced by nematode
attack on the plant.
|
|
Claim 3
A method for nematode inducible expression of a
foreign gene in a plant, comprising:
a) linking said foreign gene to a regulatory region comprising:
i) a promoter derived from a mannopine synthase gene of Agrobacterium
tumefaciens,
ii) an upstream activating sequence derived from a
mannopine synthase gene of Agrobacterium tumefaciens, and
iii) at
least one upstream activating sequence derived from an octopine synthase gene of
Agrobacterium tumefaciens;
b) inserting said foreign gene and said regulatory region in said plant,
wherein expression is induced by nematode attack on the plant.
|
|
Claim 4
A chimeric regulatory region for expressing
genes in plants comprising at least three upstream activating sequences derived
from an Agrobacterium tumefaciens octopine synthase gene operably
linked to a promoter derived from an Agrobacterium tumefaciens
mannopine synthase gene.
|
|
Claim 5
A cassette for expressing a foreign gene
comprising the foreign gene operably linked to a chimeric regulatory region
comprising at least three upstream activating sequences derived from
Agrobacterium tumefaciens octopine synthase genes operably linked to a
promoter derived from an Agrobacterium tumefaciens mannopine synthase
gene.
|
|
Claim 6
A plasmid comprising a cassette comprising a
foreign gene operably linked to a chimeric regulatory region comprising at least
three upstream activating sequences derived from Agrobacterium
tumefaciens octopine synthase genes operably linked to a promoter derived
from an Agrobacterium tumefaciens mannopine synthase gene.
|
|
Claim 7
A method of expressing a foreign gene in a
plant, comprising:
a) linking said foreign gene to a chimeric regulatory region comprising at
least three upstream activating sequences derived from an Agrobacterium
tumefaciens octopine synthase gene operably linked to a promoter derived
from an Agrobacterium tumefaciens mannopine synthase gene; and
b)
inserting said foreign gene and said chimeric regulatory region into a plant,
wherein said plant expresses said foreign gene.
|
|
Claim 8
A transgenic plant comprising a cassette
comprising a foreign gene operably linked to a chimeric regulatory region
comprising at least three upstream activating sequences derived from
Agrobacterium tumefaciens octopine synthase genes operably linked to a
promoter derived from an Agrobacterium tumefaciens mannopine synthase
gene.
|
|
Claim 9
A chimeric regulatory region for expressing
genes in plants comprising at least three upstream activating sequences derived
from an Agrobacterium tumefaciens octopine synthase gene operably
linked to an upstream activat ing sequence derived from an Agrobacterium
tumefaciens mannopine synthase gene that is operably linked to a promoter
derived from an Agrobacterium tumefaciens mannopine synthase gene.
|
|
Claim 10
A cassette for expressing a foreign gene
comprising the foreign gene operably linked to a chimeric regulatory region
comprising at least three upstream activating sequences derived from
Agrobacterium tumefaciens octopine synthase genes operably linked to an
upstream activating sequence derived from an Agrobacterium tumefaciens
mannopine synthase gene that is operably linked to a promoter derived from an
Agrobacterium tumefaciens mannopine synthase gene.
|
|
Claim 11
A method of expressing a foreign gene in a
plant, comprising:
a) linking said foreign gene to a chimeric regulatory region comprising at
least three upstream activating sequences derived from an Agrobacterium
tumefaciens octopine synthase gene operably linked to an upstream
activating sequence derived from an Agrobacterium tumefaciens
mannopine synthase gene that is operably linked to a promoter derived from an
Agrobacterium tumefaciens mannopine synthase gene; and
b)
inserting said foreign gene and said chimeric regulatory region into a plant,
wherein said plant expresses said foreign gene.
|
|
Claim 12
A transgenic plant comprising a chimeric
regulatory region for expressing genes in plants comprising at least three
upstream activating sequences derived from an Agrobacterium tumefaciens
octopine synthase gene operably linked to an upstream activating sequence
derived from an Agrobacterium tumefaciens mannopine synthase gene that
is operably linked to a promoter derived from an Agrobacterium
tumefaciens mannopine synthase gene.
|
| AU 687961 B |
|
Claim 1
A chimeric regulatory region for expressing
genes in plants comprising an upstream activating sequences derived from an
Agrobacterium tumefaciens octopine synthase gene operably linked to a
promoter derived from an Agrobacterium tumefaciens mannopine synthase
gene.
|
|
Claim 2
A cassette for expressing a gene comprising a
gene operably linked to a chimeric regulatory region comprising an upstream
activating sequences derived from an Agrobacterium tumefaciens
octopine synthase gene operably linked to a promoter derived from an
Agrobacterium tumefaciens mannopine synthase gene.
|
|
Claim 4
A chimeric regulatory region for expressing
genes in plants comprising at least two upstream activating sequences derived
from an Agrobacterium tumefaciens opine synthase gene operably linked
to a promoter derived from an Agrobacterium tumefaciens opine synthase
gene, wherein at least one of said upstream activating elements are derived from
a different opine synthase gene than said promoter.
|
|
Claim 8
A cassette for expressing a gene comprising a
gene operably linked to a chimeric regulatory region comprising at least two
upstream activating sequences derived from an Agrobacterium tumefaciens
opine synthase gene operably linked to a promoter derived from an
Agrobacterium tumefaciens opine synthase gene, wherein at least one of
said upstream activating elements are derived from a different opine synthase
gene than said promoter.
|
|
Claim 13
A cassette for inducible expression of a
foreign gene comprising said foreign gene operably linked to a regulatory region
comprising:
a) a promoter derived from a mannopine synthase gene of Agrobacterium
tumefaciens by deletion to nucleotide position -138, and
b) an
upstream activating sequence derived from a mannopine synthase gene of
Agrobacterium tumefaciens.
|
|
Claim 15
A cassette for inducible expression of a
foreign gene comprising said foreign gene operably linked to a regulatory region
comprising:
a) a promoter derived from a mannopine synthase gene of Agrobacterium
tumefaciens,
b) an upstream activating sequence derived from a
mannopine synthase gene of Agrobacterium tumefaciens, and
c) an
upstream activating sequence derived from an octopine synthase gene of
Agrobacterium tumefaciens.
|
|
Claim 16
A method for expressing a gene in a plant,
comprising the steps of:
a) linking said gene to a chimeric regulatory region comprising an upstream
activating sequence derived from an Agrobacterium tumefaciens octopine
synthase gene operably linked to a promoter derived from an Agrobacterium
tumefaciens mannopine synthase gene;
b) inserting said gene and said
chimeric regulatory region into a plant; and
c) allowing said plant to
express said gene.
|
|
Claim 17
A method for expressing a gene in a plant,
comprising the steps of:
a) linking said gene to a chimeric regulatory region comprising at least two
upstream activating sequences derived from an Agrobacterium tumefaciens
opine synthase gene operably linked to a promoter derived from an
Agrobacterium tumefaciens opine synthase gene, wherein at least one of
said upstream activating elements are derived from a different opine synthase
gene than said promoter;
b) inserting said gene and said chimeric
regulatory region into a plant; and
c) allowing said plant to express said
gene.
|
|
Claim 21
A method of inducible expression of a foreign
gene in a plant, comprising:
a) linking said foreign gene to a regulatory region comprising:
i. a promoter derived from a mannopine synthase gene of Agrobacterium
tumefaciens by deletion to nucleotide position -138, and
ii. an upstream activating sequence derived from a mannopine synthase gene of
Agrobacterium tumefaciens;
b) inserting said foreign gene and said regulatory region in said plant; and
c) inducing expression of said foreign gene.
|
|
Claim 23
A method for inducible expression of a foreign
gene in a plant, comprising:
a) linking said foreign gene to a regulatory region comprising:
i. a promoter derived from a mannopine synthase gene of Agrobacterium
tumefaciens,
ii. an upstream activating sequence derived from a mannopine synthase gene of
Agrobacterium tumefaciens; and
iii. an upstream activating sequence derived from an octopine synthase gene
of Agrobacterium tumefaciens;
b) inserting said foreign gene and said regulatory region in said plant; and
c) inducing expression of said foreign gene.
|
|
Claim 32
A chimeric regulatory region for expressing
genes in plants comprising at least three upstream activating sequences derived
from an Agrobacterium tumefaciens octopine synthase gene operably
linked to a promoter derived from an Agrobacterium tumefaciens
mannopine synthase gene.
|
|
Claim 33
A cassette for expressing a gene comprising a
gene operably linked to a chimeric regulatory region comprising at least three
upstream activating sequences derived from Agrobacterium tumefaciens
octopine synthase genes operably linked to a promoter derived from an
Agrobacterium tumefacie ns mannopine synthase gene.
|
|
Claim 34
A method of expressing a gene in a plant,
comprising the steps of:
a) linking said gene to a chimeric regulatory region comprising at least
three upstream activating sequences derived from an Agrobacterium
tumefaciens octopine synthase gene operably linked to a promoter derived
from an Agrobacterium tumefaciens mannopine synthase;
b) inserting
said gene and said chimeric regulatory region into a plant; and
c)
allowing said plant to express said gene.
|
|
Claim 37
A chimeric regulatory region for expressing a
gene in a plant comprising at least three upstream activating sequences derived
from an Agrobacterium tumefaciens opine synthase gene operably linked
to a promoter derived from an Agrobacterium tumefaciens opine synthase
gene, wherein at least one of said upstream activating elements are derived from
a different opine synthase gene than said promoter.
|
|
Claim 39
A cassette for expressing a gene in a plant
comprising a gene operably linked to a chimeric regulatory region comprising at
least three upstream activating sequences derived from Agrobacterium
tumefaciens opine synthase genes operably linked to a promoter derived from
an Agrobacterium tumefaciens opine synthase gene, wherein at least one
of said upstream activating elements are derived from a different opine synthase
gene than said promoter.
|
|
Claim 40
A chimeric regulatory region for expressing a
gene in a plant comprising at lea st three upstream activating sequences derived
from an Agrobacterium tumefaciens opine synthase gene operably linked
to a promoter derived from an Agrobacterium tumefaciens opine synthase
gene.
|
|
Claim 41
A chimeric regulatory region for expressing a
gene in a plant comprising at least two upstream activating sequences derived
from an Agrobacterium tumefaciens opine synthase gene operably linked
to a promoter derived from an Agrobacterium tumefaciens opine synthase
gene.
|
Patent applications filed by Biotechnology Research and Development
Corporation & Purdue Research Foundation
Actual pending independent claims
| EP 729514 A1 |
|
Claim 1
A chimeric regulatory region for expressing
genes in plants comprising an upst ream activating sequence derived from a first
Agrobacterium tumefaciens opine synthase gene operably linked to a promoter
derived from a second Agrobacterium tumefaciens opine synthase gene that is
different from said first Agrobacterium tumefaciens opine synthase gene.
|
|
Claim 4
A cassette for expressing a gene comprising a
gene operably linked to chimeric regulatory region comprising an upstream
activating sequence derived from a first Agrobacterium tumefaciens opine
synthase gene operably linked to a promoter derived from a second Agrobacterium
tumefaciens opine synthase gene that is different from said first Agrobacterium
tumefaciens opine synthase gene.
|
|
Claim 8
A chimeric regulatory region for expressing
genes in plants comprising at least two upstream activating sequences derived
from an Agrobacterium tumefaciens opine synthase gene operably linked to a
promoter derived from an Agrobacterium tumefaciens opine synthase gene.
|
|
Claim 12
A cassette for expressing a gene comprising a
gene operably linked to a chimeric regulatory region comprising at least two
upstream activating sequences derived from Agrobacterium tumefaciens opine
synthase genes operably linked to a promoter derived from a Agrobacterium
tumefaciens opine synthase gene.
|
|
Claim 17
A cassette for inducible expression of a
foreign gene comprising said foreign gene operably linked to a regulatory region
comprising a promoter derived from a mannopine synthase gene of Agrobacterium
tumefaciens by deletion to nucleotide position-138 and an upstream activating
sequence derived from a mannopine synthase gene of Agrobacterium tumefaciens.
|
|
Claim 19
A cassette for inducible expression of a
foreign gene comprising said foreign gene operably linked to a regulatory region
comprising a promoter derived from a mannopine synthase gene of Agrobacterium
tumefaciens, an upstream activating sequence derived from a mannopine synthase
gene of Agrobacterium tumefaciens, and an upstream activating sequence derived
from an octopine synthase gene of Agrobacterium tumefaciens.
|
|
Claim 20
A method of expressing a gene in a plant,
comprising the steps of: linking said gene to a chimeric regulatory region
comprising an upstream activating sequence derived from a first Agrobacterium
tumefaciens opine synthase gene operably linked to a promoter derived from a
second Agrobacterium tumefaciens opine synthase gene that is different from said
first Agrobacterium tumefaciens opine synthase gene; inserting said gene and
said chimeric regulatory region into a plant; and allowing said plant to express
said gene.
|
|
Claim 23
A method of expressing a gene in a plant,
comprising the steps of: linking said gene to a chimeric regulatory region
comprising at least two upstream activating sequences derived from an
Agrobacterium tumefaciens opine synthase gene operably linked to a promoter
derived from an Agrobacterium tumefaciens opine synthase; inserting said gene
and said chimeric regulatory region into a plant; and allowing said plant to
express said gene.
|
|
Claim 27
A method of inducible expressing a foreign
gene in a plant, comprising: linking said foreign gene to a regulatory region
comprising a promoter derived from a mannopine synthase gene of Agrobacterium
tumefaciens by deletion to nucleotide position-138 and an upstream activating
sequence derived from a mannopine synthase gene of Agrobacterium tumefaciens;
inserting said foreign gene and said regulatory region in said plant; and
inducing expression of said foreign gene.
|
|
Claim 29
A method for inducible expression of a foreign
gene in a plant, comprising: linking said foreign gene to a regulatory region
comprising a promoter derived from a mannopine synthase gene of Agrobacterium
tumefaciens, an upstream activating sequence derived from a mannopine synthase
gene of Agrobacterium tumefaciens, and an upstream activating sequence derived
from an octopine synthase gene of Agrobacterium tumefaciens; inserting said
foreign gene and said regulatory region in said plant; and inducing expression
of said foreign gene.
|
| CA 2174954 |
|
Claim 1
A chimeric regulatory region for expressing
genes in plants comprising an upstream activating sequence derived from an
Agrobacterium tumefaciens octopine synthase gene operably linked to a promoter
derived from an Agrobacterium tumefaciens mannopine synthase gene.
|
|
Claim 2
A cassette for expressing a gene comprising a
gene operably linked to chimeric regulatory region comprising an upstream
activating sequence derived from a Agrobacterium tumefaciens octopine synthase
gene operably linked to a promoter derived from a Agrobacterium tumefaciens
mannopine synthase gene.
|
|
Claim 4
A chimeric regulatory region for expressing
genes in plants comprising at least two upstream activating sequences derived
from an Agrobacterium tumefaciens opine synthase gene operably linked to a
promoter derived from an Agrobacterium tumefaciens opine synthase gene, wherein
at least one of said upstream activating elements are derived from a different
opine synthase gene than said promoter.
|
|
Claim 8
A cassette for expressing a gene comprising a
gene operably linked to a chimeric regulatory region comprising at least two
upstream activating sequences derived from Agrobacterium tumefaciens opine
synthase genes operably linked to a promoter derived from a Agrobacterium
tumefaciens opine synthase gene, wherein at least one of said upstream
activating elements are derived from a different opine synthase gene than said
promoter.
|
|
Claim 13
A cassette for inducible expression of a
foreign gene comprising said foreign gene operably linked to a regulatory region
comprising a promoter derived from a mannopine synthase gene of Agrobacterium
tumefaciens by deletion to nucleotide position -138 and an upstream activating
sequence derived from a mannopine synthase gene of Agrobacterium tumefaciens.
|
|
Claim 15
A cassette for inducible expression of a
foreign gene comprising said foreign gene operably linked to a regulatory region
comprising a promoter derived from a mannopine synthase gene of Agrobacterium
tumefaciens, an upstream activating sequence derived from a mannopine synthase
gene of Agrobacterium tumefaciens, and an upstream activating sequence derived
from an octopine synthase gene of Agrobacterium tumefaciens.
|
|
Claim 16
A method of expressing a gene in a plant,
comprising the steps of: linking said gene to a chimeric regulatory region
comprising an upstream activating sequence derived from a Agrobacterium
tumefaciens octopine synthase gene operably linked to a promoter derived from a
Agrobacterium tumefaciens mannopine synthase gene; inserting said gene and said
chimeric regulatory region into a plant; and allowing said plant to express said
gene.
|
|
Claim 17
A method of expressing a gene in a plant,
comprising the steps of: linking said gene to a chimeric regulatory region
comprising at least two upstream activating sequences derived from an
Agrobacterium tumefaciens opine synthase gene operably linked to a promoter
derived from an Agrobacterium tumefaciens opine synthase, wherein at least one
of said upstream activating elements are derived from a different opine synthase
gene than said promoter; inserting said gene and said chimeric regulatory region
into a plant; and allowing said plant to express said gene.
|
|
Claim 21
A method of inducible expressing a foreign
gene in a plant, comprising: linking said foreign gene to a regulatory region
comprising a promoter derived from a mannopine synthase gene of Agrobacterium
tumefaciens by deletion to nucleotide position -138 and an upstream activating
sequence derived from a mannopine synthase gene of Agrobacterium tumefaciens;
inserting said foreign gene and said regulatory region in said plant; and
inducing expression of said foreign gene.
|
|
Claim 23
A method for inducible expression of a foreign
gene in a plant, comprising: linking said foreign gene to a regulatory region
comprising a promoter derived from a mannopine synthase gene of Agrobacterium
tumefaciens, an upstream activating sequence derived from a mannopine synthase
gene of Agrobacterium tumefaciens, and an upstream activating sequence derived
from an octopine synthase gene of Agrobacterium tumefaciens; inserting said
foreign gene and said regulatory region in said plant; and inducing expression
of said foreign gene.
|
|
Claim 32
A chimeric regulatory region for expressing
genes in plants comprising at least three upstream activating sequences derived
from an Agrobacterium tumefaciens octopine synthase gene operably linked to a
promoter derived from an Agrobacterium tumefaciens mannopine synthase gene.
|
|
Claim 33
A cassette for expressing a gene comprising a
gene operably linked to a chimeric regulatory region comprising at least three
upstream activating sequences derived from Agrobacterium tumefaciens octopine
synthase genes operably linked to a promoter derived from a Agrobacterium
tumefaciens mannopine synthase gene.
|
|
Claim 34
A method of expressing a gene in a plant,
comprising the steps of: linking said gene to a chimeric regulatory region
comprising at least three upstream activating sequences derived from an
Agrobacterium tumefaciens octopinesynthase gene operably linked to a promoter
derived from an Agrobacterium tumefaciens mannopine synthase; inserting said
gene and said chimeric regulatory region into a plant; and allowing said plant
to express said gene.
|
Actual granted independent claims
Patents granted to Monsanto
|
US
5352605
|
|
Claim 1
A chimeric gene which is expressed in plant
cells comprising:
(i) a promoter from a cauliflower mosaic virus, said promoter selected from
the group consisting of a CaMV (35S) promoter isolated from CaMV
protein-encoding DNA sequences and a CaMV (19S) promoter isolated from CaMV
protein-encoding DNA sequences, and
(ii) a structural sequence which is
heterologous with respect to the promoter.
|
|
Claim 4
A plant cell which comprises a chimeric gene
that contains:
(i) a promoter from cauliflower mosaic virus, said promoter selected from the
group consisting of a CaMV (35S) promoter and a CaMV (19S) promoter, wherein
said promoter is isolated from CaMV protein-encoding DNA sequences, and
(ii) a structural sequence which is heterologous with respect to the promoter.
|
|
Claim 7
An intermediate plant transformation plasmid
which comprises:
(i) a region of homology to an Agrobacterium tumefaciens vector,
(ii) a T-DNA border region from Agrobacterium tumefaciens
and
(iii) a chimeric gene, wherein the chimeric gene is located between the
T-DNA border and the region of homology,
said chimeric gene comprising:
(i) a promoter from cauliflower mosaic virus, said promoter selected from the
group consisting of a CaMV(35S) promoter and a CaMV(19S) promoter, and
(ii) a structural sequence which is heterologous with respect to the promoter.
|
|
Claim 8
A plant transformation vector which comprises:
(i) a disarmed plant tumor inducing plasmid of Agrobacterium tume
faciens and
(ii) a chimeric gene, wherein the chimeric gene contains a
promoter from cauliflower mosaic virus, said promoter selected from the group
consisting of a CaMV(35S) promoter and a CaMV(19S) promoter, and
(iii) a
structural sequence which is heterologous with respect to the promoter.
|
|
Claim 13
A DNA construct comprising:
(i) a CaMV promoter selected from the group consisting of
(a) a CaMV 35S promoter isolated from CaMV protein-encoding DNA sequences and
(b) a CaMV 19S promoter isolated from CaMV protein-encoding DNA sequences,
and
(ii) a DNA sequence of interest heterologous to (i), wherein (ii) is under
the regulatory control of (i) when said construct is transcribed in a plant
cell.
|
|
Claim 14
A chimeric gene which is transcribed and
translated in plant cells, said chimeric gene comprising:
(i) a promoter from cauliflower mosaic virus, said promoter selected from the
group consisting of:
(a) a CaMV 35S promoter region free of CaMV protein-encoding DNA sequences
and
(b) a CaMV 19S promoter region free of CaMV protein-encoding DNA
sequences, and
(ii) a DNA sequence which is heterologous with respect to the promoter.
|
|
Claim 15
A chimeric gene which is expressed in plants
cells comprising:
(i) a promoter from a cauliflower mosaic virus, said promoter selected from
the group consisting of
(a) a CaMV(35S) promoter region free of CaMV protein-encoding DNA sequences
and
(b) a CaMV(19S) promoter region free of CaMV protein-encoding DNA
sequences, and
(ii) a DNA sequence which is heterologous with respect to the promoter.
|
|
Claim 16
A chimeric gene which is transcribed in plants
cells comprising:
(i) a promoter from a cauliflower mosaic virus, said promoter selected from
the group consisting of
(a) a CaMV(35S) promoter free of CaMV protein-encoding DNA sequences and
(b) a CaMV(19S) promoter free of CaMV protein-encoding DNA sequences,
(ii) a DNA sequence which is heterologous with respect to the promoter and
(iii) a 3' non-translated polyadenylation signal sequence.
|
|
Claim 17
A plant cell which comprises a chimeric gene
where said chimeric gene comprises:
(i) a promoter from cauliflower mosaic virus, said promoter selected from the
group consisting of
a CaMV(35S) promoter and a CaMV(19S) promoter, wherein
said promoter is free of CaMV protein-encoding DNA sequences, and
(ii) a
DNA sequence which is heterologous with respect to the promoter and
(iii) a
3' non-translated polyadenylation signal sequence.
|
Patents on entire 35S and 19S promoters granted to Monsanto
Actual granted independent claims
|
US
5530196
|
|
Claim 1
A differentiated dicotyledonous plant
comprising plant cells containing a chime ric gene which comprises:
a) a promoter from cauliflower mosaic virus, said promoter selected from the
group consisting of
- a CaMV(35S) promoter free of CaMV protein-encoding DNA sequences and
- a CaMV(19S) promoter free of protein-encoding DNA sequences, and
b) a structural sequence which is heterologous with respect to the promoter.
|
|
Claim 4
A differentiated dicotyledonous plant
comprising plant cells containing in the plant genome a chimeric gene which
comprises:
- a promoter from cauliflower mosaic virus, said promoter selected from the
group consisting of a CaMV(35S) promoter and a CaMV(19S) promoter, and
- a DNA sequence which is heterologous with respect to the promoter.
|
|
Claim 5
A differentiated dicotyledonous plant
regenerated from plant cells, said plant cells containing a chimeric gene which
comprises:
i. a promoter from cauliflower mosaic virus, said promoter selected from the
group consisting of a CaMV(35S) promoter and a CaMV(19S) promoter, and
ii. a DNA sequence which is heterologous with respect to the promoter.
|
|
US
5858742
|
|
Claim 1
A method for transforming a plant cell which
comprises transforming a plant cell with a chimeric DNA construct containing:
a) a promoter isolated from cauliflower mosaic virus (CaMV), said promoter
selected from the group consisting of
- a CaMV(19S) promoter derived from the CaMV(19S) gene and
- a CaMV(35S) promoter derived from the CaMV(35S) gene, and
b) a DNA sequence which is heterologous with respect to the promoter;
wherein the promoter regulates the transcription of the DNA sequence.
|
|
US
6255560
|
|
Claim 1
A chimeric gene which is expressed in plant
cells comprising:
a) a promoter from cauliflower mosaic virus (CaMV), wherein said promoter is
the CaMV(19S) promoter or the CaMV(35S) promoter, operably linked to
b) a
DNA sequence which is heterologous with respect to the promoter, wherein:
- the promoter regulates the transcription of the DNA sequence, and
- the DNA sequence encodes a polypeptide conferring increased antibiotic
resistance to a plant or plant cell containing the DNA sequence relative to a
wild-type plant or plant cell.
|
|
Claim 3
A plant cell comprising a chimeric gene which
comprises:
a) a promoter from cauliflower mosaic virus (CaMV), wherein said promoter is
the CaMV(19S) promoter or the CaMV(35S) promoter, operably linked to
b) a
DNA sequence which is heterologous with respect to the promoter, wherein:
- the promoter regulates the transcription of the DNA sequence, and
- the DNA sequence encodes a polypeptide conferring increased antibiotic
resistance to the plant cell relative to a wild-type plant cell.
|
|
Claim 6
An intermediate plant transformation plasmid
which comprises:
a) a region of homology to an A. tumefaciens vector;
b) a T-DNA
border from A. tumefaciens, and
c) a chimeric gene,
wherein the chimeric gene is located between the T-DNA border and the region
of homology, said chimeric gene comprising a promoter from cauliflower mosaic
virus (CaMV), wherein said promoter is the CaMV(19S) promoter or the CaMV(35S)
promoter, operably linked to a DNA sequence which is heterologous with respect
to the promoter, wherein:
a) the promoter regulates the transcription of the DNA sequence, and
b)
the DNA sequence encodes a polypeptide conferring increased antibiotic
resistance to a plant or plant cell containing the DNA sequence relative to a
wild-type plant or plant cell.
|
|
Claim 9
A plant transformation vector which comprises a
modified plant tumor inducing plasmid of A. tumefaciens which is
capable of inserting a chimeric gene into susceptible plant cells, wherein the
chimeric gene comprises a promoter from cauliflower mosaic virus (CaMV), wherein
said promoter is the CaMV(19S) pr omoter or the CaMV(35S) promoter, operably
linked to a DNA sequence which is heterologous with respect to the promoter,
wherein:
a) the promoter regulates the transcription of the DNA sequence, and
b) the DNA sequence encodes a polypeptide conferring increased antibiotic
resistance to a plant or plant cell containing the DNA sequence relative to a
wild-type plant or plant cell.
|
|
Claim 12
A differentiated dicotyledonous plant
comprising plant cells containing a chimeric gene which comprises a promoter
from cauliflower mosaic virus (CaMV), wherein said promoter is the CaMV(19S)
promoter or the CaMV(35S) promoter, operably linked to a DNA sequence encoding
said polypeptide which is heterologous with respect to the promoter, wherein:
a) the promoter regulates the transcription of the DNA sequence, and
b)
the DNA sequence encodes a polypeptide conferring increased antibiotic
resistance to the plant relative to a wild-type plant.
|
|
EP
131623 B2
|
|
Claim 1
A chimeric gene capable of expressing a
neomycin phosphotransferase polypeptide in plant cell conferring antibiotic
resistance to the plant when inserted into the plant genome, comprising in
sequence:
a) a promoter region from a ribulose-1,5-bis-phosphate carboxylase small
subunit gene;
b) a 5' non-translated region;
c)a structural coding sequence encoding neomycin phosphotransferase I oy II;
and
d) a 3' non-translated region of a gene naturally expressed in plant cells,
said region encoding a signal sequence for polyadnylation of mRNA; said promoter
being heterologous with respect to the structural coding sequence.
Claim 4
A chimeric gene capable of expressing a
polypeptide in plant cells comprising in sequence:
a) a full-length transcript promoter region isolated from cauliflower mosaic
virus
b) a 5' non-translated region
c) a structural coding
sequence
d) a 3' non-translated region of a gene naturally expressed in
plants,
said region encoding a signal sequence for polyadenylation of
mRNA,
said structural coding sequence being heterologous with respect to
said promoter region.
|
Patents granted to the Rockefeller University
Actual granted independent claims
|
US
5097025
|
|
Claim 1
An isolated DNA segment consisting of the
nucleotide sequence:
5'-CGACCAGCAT CGTGGAAAAA GAAGACGTTC CAACCACGTC
TTCAAAGC-3'*.
*sequence of subdomain B2 of 35S CaMV promoter.
|
|
Claim 2
A DNA sequence consisting of:
a) a first
nucleotide sequence 5'-CGAGGAGCAT CGTGGAAAAA GAAGACGTTC CAACCACGTC TTCAAAGC-3'*
and
b) a second nucleotide sequence corresponding to domain A of the CaMV
35S promoter coupled to said first nucleotide sequence by means of a synthetic
multilinker.
*sequence of subdomain B2 of 35S CaMV promoter.
|
|
Claim 3
A DNA sequence consisting of:
a) a first
nucleotide sequence 5'-CATCGTTGAAG ATGCCTCTGC CGACAGTGGT CCCAAAGATG GACCCCCACC
CAC-3'* and
b) a second nucleotide sequence corresponding to domain A of
the CaMV35S promoter coupled to said first nucleotide sequence by means of a
synthetic multilinker.
*sequence of subdomain B3 of 35S CaMV promoter.
|
|
Claim 4
A DNA sequence consisting of:
a) a first
nucleotide sequence 5'-ATTCC ATTGCCC AGCTATCTGT CACTTTATTG TGAAGATAGT GGAAAAGGAA
GGTGGCTCCT ACAAATGCCA TCATTGCGAT AAAGGAAAGG CC-3'* and
b) a second
nucleotide sequence corresponding to domain A of the CaMV 35S promoter coupled
to said first nucleotide sequence by means of a synthetic multilinker.
*sequence of subdomain B4 of 35S CaMV promoter.
|
|
Claim 5
A DNA sequence consisting of:
a) a first
nucleotide sequence 5'-TGAGACTTTT CAACAAAGGG TAATATCCGG AAACCTCCTC GGATT-3'*
and
b) a second nucleotide sequence corresponding to domain A of the CaMV
35S promoter coupled to said first nucleotide sequence by means of a synthetic
multilinker.
*sequence of subdomain B5 of 35S CaMV 35S promoter.
|
|
Claim 6
An isolated DNA segment consisting of the
nucleotide sequence:
5'-CATCGTTGAAG ATGCCTCTGC CGACAGTGGT CCCAAAGATG
GACCCCCACC CAC-3'*.
*sequence of subdomain B3 of 35S CaMV promoter.
|
|
Claim 7
An isolated DNA segment consisting of the
nucleotide sequence:
5'-ATTCC ATTGCCC AGCTATCTGT CACTTTATTG TGAAGATAGT
GGAAAAGGAA GGTGGCTCCT ACAAATGCCA TCATTGCGAT AAAGGAAAGG CC-3'*.
*sequence of subdomain B4 of 35S CaMV promoter.
|
|
Claim 8
An isolated DNA segment consisting of the
nucleotide sequence:
5'-TGAGACTTTT CAACAAAGGG TAATATCCGG AAACCTCCTC
GGATT-3'*.
*sequence of subdomain B5 of 35S CaMV promoter.
|
|
Claim 9
A DNA sequence consisting of:
a) a first
nucleotide sequence 5'-CGAGGAGCAT CGTGGAAAAA GAAGACGTTC CAACCACGTC TTCAAAGC-3'*
and
b) a second nucleotide sequence corresponding to the minimal promoter
region of the CaMV 35S promoter coupled to said first nucleotide sequence by
means of a synthetic multilinker.
*sequence of subdomain B2 of 35S CaMV promoter.
|
|
Claim 10
A DNA sequence consisting of:
a) a first
nucleotide sequence 5'-CATCGTTGAAG ATGCCTCTGC CGACAGTGGT CCCAAAGATG GACCCCCACC
CAC-3'*
and
b) a second nucleotide sequence corresponding to the
minimal promoter region of the CaMV 35S promoter coupled to said first
nucleotide sequence by means of a s ynthetic multilinker.
*sequence of subdomain B3 of CaMV 35S promoter
|
|
Claim 11
A DNA sequence consisting of:
a) a first
nucleotide sequence 5'-ATTCC ATTGCCC AGCTATCTGT CACTTTATTG TGAAGATAGT GGAAAAGGAA
GGTGGCTCCT ACAAATGCCA TCATTGCGAT AAAGGAAAGG CC-3'* and
b) a second
nucleotide sequence corresponding to the minimal promoter region of the CaMV 35S
promoter coupled to said first nucleotide sequence by means of a synthetic
multilinker.
*sequence of subdomain B4 of 35S CaMV promoter
|
|
Claim 12
A DNA sequence consisting of:
a) a first
nucleotide sequence 5'-TGAGACTTTT CAACAAAGGG TAATATCCGG AAACCTCCTC GGATT-3'*
and
b) a second nucleotide sequence corresponding to the minimal promoter
region of the CaMV 35S promoter coupled to said first nucleotide sequence by
means of a synthetic multilinker.
*sequence of subdomain B5 of 35S CaMV promoter
|
Patent granted to the Rockefeller University
Actual granted independent claims
|
US
5110732
|
|
Claim 1
In a method for the expression of a chimeric
plant gene, the improvement which comprises the use of a tissue-specific
promoter fragment which causes tissue-specific expression in leaves, stems,
cotyledons, and vascular tissue of the hypocotyl while causing detectable levels
of expression in root vascular tissue when operably coupled directly to a DNA
segment corresponding to the -72 to +8 promoter fragment of the Cauliflower
Mosaic Virus 35S gene, said tissue-specific promoter fragment having the
sequence:
5'-TGAGACTTTT CAACAAAGGG TAATATCCGG AAACCTCCTC GGATTCCATT GCCCAGCTAT
CTGTCACTTT
ATTGTGAAGA TAGTGGAAAA GGAAGGTGGC TCCTACAAAT GCCATCATTG
CGATAAAGGA AAGGCCATCG
TTGAAGATGC CTCTGCCGAC AGTGGTCCCA AAGATGGACC
CCCACCCCAC GAGGAGCATC GTGGAAAAAG
AAGACGTTCC AACCACGTCT TCAAAGCAAG
TGGATTGATG TGATA-3'*
*sequence corresponds to the complete domain B -from -343 to -90 nucleotides-
of the 35S CaMV promoter
|
|
Claim 2
A chimeric plant gene comprising in sequence in
the 5' to 3' direction a tissue-specific promoter fragment consisting
essentially of the sequence:
5'-TGAGACTTTT CAACAAAGGG TAATATCCGG AAACCTCCTC GGATTCCATT GCCCAGCTAT
CTGTCACTTT
ATTGTGAAGA TAGTGGAAAA GGAAGGTGGC TCCTACAAAT GCCATCATTG
CGATAAAGGA AAGGCCATCG
TTGAAGATGC CTCTGCCGAC AGTGGTCCCA AAGATGGACC
CCCACCCCAC GAGGAGCATC GTGGAAAAAG
AAGACGTTCC AACCACGTCT TCAAAGCAAG
TGGATTGATG TGATA-3'*,
operably coupled directly to the -72 to +8 promoter fragment of the CaMV 35S
gene, said -72 to +8 promoter fragment operably coupled to a structural gene.
*sequence corresponds to the complete domain B from -343 to -90 nucleotides
of the 35S CaMV promoter.
|
|
Claim 5
A tissue-specific promoter fragment which
functions in plants to cause tissue-specific expression in the leaves, stems,
cotyledons and the vascular tissue of the hypocotyl and detachable levels of
expression in root vascular tissue operably coupled directly to a DNA segment
corresponding to the -72 to +8 promoter fragment of the Cauliflower Mosaic Virus
35S gene, said tissue-specific promoter fragment having the sequence from its 5'
to 3' termini:
5'-TGAGACTTTT CAACAAAGGG TAATATCCGG AAACCTCCTC GGATTCCATT GCCCAGCTAT
CTGTCACTTT
ATTGTGAAGA TAGTGGAAAA GGAAGGTGGC TCCTACAAAT GCCATCATTG
CGATAAAGGA AAGGCCATCG
TTGAAGATGC CTCTGCCGAC AGTGGTCCCA AAGATGGACC
CCCACCCCAC GAGGAGCATC GTGGAAAAAG
AAGACGTTCC AACCACGTCT TCAAAGCAAG
TGGATTGATG TGATA-3'*
*sequence corresponds to the complete domain B -from -343 to -90 nucleotides-
of the 35S CaMV promoter.
|
Patent application filed by Monsanto
Actual pending independent claims
| EP 1210446 |
|
Claim 1
A DNA sequence comprising an ubiquitin
regulatory system lacking heatshock elements.
|
|
Claim 2
A DNA sequence comprising an ubiquitin
regulatory system that is not heat inducible.
|
Patents granted to Mycogen
Actual granted independent claims
|
US
5510474
|
|
Claim 1
An isolated DNA fragment, useful in effecting
expression in both monocots and dicots of coding sequences placed 3' to said
fragment, wherein said DNA is approximately 2 kb in length, and said fragment
further comprises, in the following order beginning with the 5' most element and
proceeding toward the 3' terminus of said DNA fragment:
a) two heat shock elements, which overlap;
b) a promoter comprising a
transcription start site;
c) an intron of about 1 kb in length;
d) and
a translation start site;
wherein said DNA fragment comprising said
elements (i)-(iv) regulates gene expression in both dicots and monocots,
and
wherein DNA fragment comprises the nucleotide sequence shown from
position -899 to 1092 of the maize ubiquitin sequence listed in FIG. 2.
|
|
US
5614399
|
|
Claim 1
A method for selective heat shock induced
enhancement of the constitutive expression of a structural gene in a plant cell
comprising the steps of:
a) transforming said plant cell with a DNA construct comprising an
approximately 2 kb plant ubiquitin regulatory region operably joined to a DNA
sequence of interest, wherein said plant ubiquitin regulatory region is from a
plant ubiquitin gene and comprises:
- at least one heat shock element,
- a promoter,
- a transcription start site, and
- an intron; and
b) selectively applying stress conditions of high temperature to said
transformed plant cell thereby inducing enhancement in expression of said DNA
sequence of interest.
|
|
US
6020190
|
|
Claim 1
A DNA construct comprising:
a) a DNA sequence no larger than 2 kb, said DNA sequence comprising a plant
ubiquitin regulatory system, wherein said regulatory system contains:
- a heat shock element, and
- an intron, said intron being located at 3' to said heat shock element, and
b) a plant-expressible structural gene wherein said structural gene is placed
under the regulatory control of said plant ubiquitin regulatory system.
|
|
US
6054574
|
|
Claim 1
A DNA fragment of approximately 2 kb in length,
said DNA fragment comprising a plant ubiquitin regulatory system, wherein said
regulatory system contains:
a) a promoter comprising a transcription start site,
b) one or more heat
shock elements positioned 5' to said transcription start site, and
c) an
intron positioned 3' to said transcription start site,
wherein said
regulatory system is capable of regulating constitutive and inducible gene
expression in both dicots and monocots such that the level of said constitutive
gene expression in monocots is about one-third that obtained in said inducible
gene expression in monocots.
|
|
Claim 9
A recombinant DNA construct comprising:
a) a DNA fragment of approximately 2 kb in length, said DNA fragment
comprising a plant ubiquitin regulatory system, wherein said plant ubiquitin
regulatory system contains:
- a promoter comprising a transcription start site,
- one or more heat shock elements positioned 5' to said transcription start
site,
- a translational start site, and
an intron positioned 3' to said
transcription start site and 5' to said translational start site,
wherein
said plant ubiquitin regulatory system is capable of regulating constitutive and
inducible gene expression in both dicots and monocots such that said
constitutive gene expression in monocots is at a level about one-third that
obtained in said inducible gene expression in monocots, and
b) a plant-expressible heterologous structural gene positioned 3' to said
plant ubiquitin regulatory system and
c) a polyadenylation signal
positioned 3' to said structural gene,
wherein said heterologous gene is
placed under the regulatory control of said plant ubiquitin regulatory system.
|
|
Claim 18
A DNA fragment, useful in effecting expression
in both monocots and dicots of coding sequences placed 3' to said fragment,
wherein said DNA is isolated or incorporated into a larger piece of DNA but in a
position other than in the 5' sequence of a plant ubiquitin gene, is
approximately 2 kb in length, and said DNA fragment further comprises, in the
following order beginning with the 5' most element and proceeding toward the 3'
terminus of said DNA fragment:
(a) one or more heat shock elements, which elements may or may not be
overlapping;
(b) a promoter comprising a transcription start site; and
(c) an intron of about 1 kb in length;
and wherein said DNA fragment
comprising said elements (a)-(c) is capable of regulating gene expression in
both dicots and monocots.
wherein said DNA is isolated or incorporated into a larger piece
of DNA but in a position other than in the 5' sequence of a plant ubiquitin
gene, meaning of this?? Does not regulate the plant ubiquitin
gene, is it that? or is not related to the 5'sequence of a plant ubiquitin gene
at all??
|
| CA 1339684 |
|
Claim 1
A DNA fragment approximately 2kb in length,
said DNA fragment comprising a plant ubiquitin regulatory system, wherein said
regulatory system contains:
a) overlapping heat shock elements and
b) an intron, and
wherein said regulatory system is capable of regulating
constitutive and inducible gene expression in both dicots and monocots.
|
|
Claim 11
A recombinant DNA construct comprising:
a) a DNA fragment approximately 2kb in length, said DNA fragment comprising a
plant ubiquitin regulatory system, wherein said regulatory system contains:
- overlapping heat shock elements and
- an intron; and
wherein said regulatory system is capable of regulating
constitutive and inducible gene expression in both dicots and monocots, and
b) a plant-expressible heterologous structural gene wherein said heterologous
structural gene is placed under the regulatory control of said plant ubiquitin
regulatory system.
|
|
Claim 23
A method for the constitutive expression of a
structural gene and the selected stress-induced enhancement in expression of
said structural gene in a plant cell comprising the steps of:
a)
transforming said plant cell with a DNA construct comprising an approximately
2kb plant ubiquitin regulatory system, wherein is found a heat shock element and
an intron, and
b) a plant-expressible structural gene that is under the
regulatory control of said plant regulatory system, and
c) selectively
applying stress conditions of high temperature to said transformed plant cell
thereby inducing enhancement in expression of said structural gene.
|
|
EP
342926 B1
|
|
Claim 1
A DNA sequence no larger than 2 kb, said DNA
sequence comprising a plant ubiquitin regulatory system, wherein said regulatory
system contains:
a) a heat shock element and
b) an intron.
|
|
Claim 10
A DNA construct comprising:
a) a DNA sequence no larger than 2 kb, said DNA sequence comprising a plant
ubiquitin regulatory system, wherein said regulatory system contains:
- a heat shock element, and
- an intron; and
b) a plant-expressible structural gene wherein said structural gene is placed
under the regulatory control of said plant ubiquitin regulatory system.
|
|
Claim 12
A method for the constitutive expression of a
structural gene and the selected stress-induced enhancement in expression of
said structural gene in a plant cell comprising the steps of:
a) transforming said plant cell with a DNA construct comprising:
i. a plant ubiquitin regulatory system, wherein is found a heat shock element
and an intron, and
ii. a plant-expressible structural gene that is under the regulatory control
of said plant ubiquitin regulatory system, and
b) selectively applying stress conditions to said transformed plant cell
thereby inducing enhancement in expression of said structural gene.
|
1. Patents on the ARE of maize ADH and maize aldolase promoters
Actual granted independent claims
Patent granted to Lubrizol Enterprises and CSIRO
| CA 1338858 |
|
Claim 1
A recombinant DNA molecule comprising: (a) an
anaerobic regulatory element; (b) a plant-expressible promoter located 3' to
said anaerobic regulatory element, and (c) a plant expressible structural gene
located 3' to said plant-expressible promoter such that said structural gene is
placed under the regulatory control of said promoter and said anaerobic
regulatory element, wherein said structural gene is not in nature under the
regulatory control of said anaerobic regulatory element.
|
|
Claim 25
A method for selective expression of a
plant-expressible structural gene under anaerobic conditions in plant tissue
which comprises the steps of:
a) constructing a recombinant DNA molecule which comprises
i. an anaerobic regulatory control element;
ii. a plant-expressible promoter located 3' to said anaerobic regulatory
element, and
iii. a plant expressible structural gene located 3' to said plant express
ible promoter such that said structural gene is placed under the regulatory
control of said promoter and said anaerobic regulatory element;
b) transforming said plant tissue with said recombinant DNA molecule, and
c) placing said transformed plant cell under anaerobic conditions so that
said plant-expressible structural gene is expressed.
|
Patent granted to Mycogen Plant Science Inc. and CSIRO
|
EP
278658 B1
|
|
Claim 1
A method for selective expression of a
plant-expressible structural gene under anaerobic conditions in plant tissue,
which method comprises using as an anaerobic regulatory element a recombinant
DNA molecule comprising a sequence selected from:
1) 5'-GCTGGTTTCT-3'
2) 5'-CGTGGTTTGCTTGCC-3', or a sequence having about 66% or greater homology
thereto
3) 5'-CGAGCCTTTCTTCCC-3'
4) 5'-CTGCCTCCCTGGTTTCT-3', and
5) 5'-CTGCAGCCCCGGTTTCG-3', or a sequence having about 66% or greater homology
thereto,
a plant-expressible promoter being located 3' to said anaerobic
regulatory element, and a plant-expressible structural gene being located 3' to
said plant-expressible promoter such that said structural gene is placed under
the regulatory control of said promoter and said anaerobic regulatory element.
|
Patent granted to Mycogen Plant Science Inc. and Lubrizol
Enterprises
|
US
5001060
|
|
Claim 1
A recombinant DNA molecule comprising:
(a) an anaerobic regulatory element;
(b) a plant-expressible promoter
located 3' to said anaerobic regulatory element, and
(c) a
plant-expressible structural gene located 3' to said plant-expressible
promoter
such that said structural gene is placed under the regulatory
control of said promoter and said anaerobic regulatory element
wherein
said structural gene is not in nature under the regulatory control of said
anaerobic regulatory element.
|
2. Patents on the recombinant promoter EMU
Patent granted to Lubrizol Genetics Inc.
| AU 643521 B2 |
|
Claim 1
A recombinant promoter molecule for enhancing
expression of a plant-expressible structural gene in a monocot plant cell
comprising:
a) a plurality of enhancer elements selected from the group consisting of the
ARE and the OCS elements;
b) a truncated, plant expressible promoter
providing a TATA box region necessary to initiate transcription positioned 3' to
said plurality of enhancer elements; and
c) a nucleotide sequence naturally
found as an intron positioned between the transcription start site and the
translation start site in a plant-expressible gene;
whereby a
plant-expressible structural gene placed 3' to said recombinant promoter
molecule is expressed in said monocot plant cell under regulatory control of
said recombinant promoter molecule.
|
Patent granted to Mycogen Plant Science Inc. and CSIRO
|
EP
459643 B1
|
|
Claim 1
A recombinant promoter molecule for enhancing
expression of a plant-expressible structural gene in a monocot plant cell
comprising:
a) a plurality of ARE enhancer elements;
b) a truncated, plant
expressible promoter providing a TATA box region necessary to initiate
transcription positioned 3' to said plurality of enhancer elements; and
c)
a nucleotide sequence naturally found as an intron positioned between the
transcription start site and the translational start site in a plant-expre
ssible gene;
whereby a plant-expressible structural gene placed 3' to said
recombinant promoter molecule is expressed in said monocot plant cell under
regulatory control of said recombinant promoter molecule.
|
Patent granted to CSIRO
|
US
5290924
|
|
Claim 1
A recombinant promoter molecule, useful for
enhancing expression of a plant-expressible structural gene in a monocot plant
cell, said promoter molecule comprising:
a) a plurality of enhancer elements selected from the group consisting of
only ARE elements, only OCS elements, and combinations of ARE and OCS elements;
b) a truncated, plant expressible promoter, providing a TATA box region
and a transcription start site, said promoter selected from the group consisting
of .DELTA.35S and .DELTA.ADH positioned 3' to said plurality of enhancer
elements wherein said truncated promoter excludes the presence of enhancer
sequences and wherein said truncated promoter is recombined with said plurality
of enhancer elements positioned 5' to said truncated promoter; and
c) a
maize Adh1 intron positioned 3' to said transcription start site whereby a
plant-expressible structural gene, placed 3' to said recombinant promoter
molecule, is expressed in said monocot plant cell under regulatory control of
said recombinant promoter molecule.
|
Patents granted to Pioneer Hi-Bred International Inc.
Actual granted independent claims
|
US
6072050
|
|
Claim 1
A synthetic DNA promoter sequence functional in
a plant cell, said promoter sequence comprising:
a TATA motif,
a transcription start site, and
a region between said
TATA motif and said start site that is at least 64% GC-rich;
wherein said
region is not a region between a TATA motif and a transcription start site of
native maize ubiquitin promoter, and
wherein said promoter sequence is set
forth in SEQ ID NO:10.
|
|
Claim 2
A synthetic DNA promoter sequence functional in
a plant cell, said promoter sequence comprising:
a TATA motif,
a transcription start site, and
a region between
said TATA motif and said start site that is at least 64% GC-rich;
wherein
said region is not a region between a TATA motif and a transcription start site
of native maize ubiquitin promoter, and
wherein said promoter sequence is
set forth in SEQ ID NO:1.
|
|
Claim 3
An expression cassette comprising
a synthetic promoter comprising:
a TATA motif,
a transcription
start site and
a region between said TATA motif and said start site that is
at least 64% GC rich,
a structural gene operatively linked to said
promoter, and
a transcription end site polyadenylation signal;
wherein said region is not a region between a TATA motif and a transcription
start site of native maize ubiquitin promoter, and
wherein sequence of
said promoter is set forth in SEQ ID NO:1.
|
|
Claim 4
An expression cassette comprising
a synthetic promoter comprising:
a TATA motif,
a transcription
start site and
a region between said TATA motif and said start site that is
at least 64% GC rich,
a structural gene operatively linked to said
promoter, and
a transcription end site polyadenylation signal;
wherein said region is not a region between a TATA motif and a transcription
start site of native maize ubiquitin promoter, and
wherein sequence of said
promoter is set forth in SEQ ID NO:10.
|
|
Claim 5
An expression cassette comprising
a synthetic promoter comprising:
a TATA motif,
a transcription
start site and
a region between said TATA motif and said start site that
is at least 64% GC rich,
a structural gene operatively linked to said
promoter,
a transcription end site polyadenylation signal, and
an
upstream element operatively linked to said promoter so that transcription is
enhanced;
wherein said region is not a region between a TATA motif and a
transcription start site of native maize ubiquitin promoter; and
wherein
sequence of said upstream element is set forth in SEQ ID NO:2.
|
|
Claim 7
A synthetic upstream element having a sequence
set forth in SEQ ID NO:2.
|
|
Claim 8
An expression cassette comprising:
a promoter sequence;
a structural gene operatively linked to said
promoter sequence;
a polyadenylation signal; and
a synthetic upstream
element comprising SEQ ID NO:2 operatively linked to said promoter so that
expression is enhanced.
|
|
Claim 13
An isolated nucleotide sequence comprising a
DNA enhancer sequence comprising the nucleotide sequence set forth in SEQ ID No:
5.
|
|
Claim 14
A nucleotide sequence comprising a promoter
construct,
said construct comprising in operable linkage a core promoter
sequence and a Ubi-1 UAR,
wherein said Ubi-1 UAR is a maize Ubi UAR
comprising the sequence set forth in SEQ ID No: 13.
|
|
Claim 15
An expression cassette comprising in operable
linkage:
a core promoter sequence,
a Ubi UAR operably linked upstream to said
core promoter to form a synthetic promoter construct,
a nucleotide sequence
of interest operably linked to said synthetic promoter, and
a
polyadenylation signal;
wherein said Ubi-1 UAR comprises the sequence set
forth in SEQ ID No:13.
|
|
AU
729929 B2
|
|
Claim 1
A synthetic DNA plant promoter sequence, said
sequence comprising:
a TATA motif,
a transcription start site, and
a region between said
TATA motif and said start site that is at least 64% GC-rich.
|
|
Claim 3
An expression cassette comprising
a synthetic promoter comprising
a TATA motif,
a transcription start
site and
a region there between that is at least 65% GC rich,
a
structural gene operatively linked to said promoter, and
a transcription
end site polyadenylation signal.
|
|
Claim 17
A synthetic upstream element having a sequence
of SEQ ID NO:2.
|
|
Claim 18
An expression cassette comprising:
a
promoter sequence;
a structural gene operatively linked to said promoter
sequence;
a polyadenylation signal; and
a synthetic upstream element
comprising SEQ ID NO:2 operatively linked to said promoter sequence so that
expression is enhanced.
|
|
Claim 23
A synthetic DNA plant promoter sequence
functional in a plant cell, said sequence comprising:
a TATA motif,
a transcription start site, and
a region between said
TATA motif and said start site that is at least 64% GC-rich; wherein said
promoter sequence is less than 1000 bp.
|
| EP 1056875 A1 |
|
Claim 1
A synthetic DNA plant promoter sequence, said
sequence comprising:
a TATA motif ;
a transcription start site; and
a region between
said TATA motif and said start site that is at least about 64% GC-rich.
|
|
Claim 4
An expression cassette comprising:
a synthetic promoter comprising:
a TATA motif ;
a transcription
start site and a region there between that is at least about 64% GC rich;
a structural gene operatively linked to said promoter; and
a transcription
end site polyadenylation signal.
|
|
Claim 19
A synthetic upstream element having a sequence
of SEQ ID NO: 2.
|
|
Claim 20
An expression cassette comprising:
a promoter sequence;
a structural gene operatively linked to said
promoter sequence;
a polyadenylation signal; and
a synthetic upstream
element homologous to SEQ ID NO: 2 operatively linked to said promoter so that
expression is enhanced.
|
|
Claim 25
A DNA sequence comprising:
a promoter construct, said construct comprising in operable linkage:
a
core synthetic promoter sequence comprising a TATA motif,
a transcription
start site, and
a region between said TATA motif and said start site that
is at least 64% GC-rich; and
an upstream activating region operably linked
to said core synthetic promoter.
|
|
Claim 32
An expression cassette comprising:
a core synthetic promoter sequence comprising a TATA motif,
a
transcription start site, and
a region between said TATA motif and said
start site that is at least 64% GC-rich;
an upstream activating region
operably linked to said synthetic promoter to enhance transcription;
a
structural gene operably linked to said synthetic promoter; and
a
polyadenylation signal.
|
|
Claim 38
A method for controlling the level of
expression of a transgenic nucleotide sequence in a plant cell said method
comprising transforming with an expression cassette comprising a promoter having
at least one Ubi-1 UAR.
|
|
Claim 40
A nucleotide sequence comprising:
a promoter construct, said construct comprising in operable linkage:
a
core promoter sequence; and
a Ubi-I UAR.
|
|
Claim 46
An expression cassette comprising in operable
linkage:
a core promoter sequence;
a Ubi UAR operably linked to said core
promoter to form a synthetic promoter construct;
a nucleotide sequence of
interest operably linked to said synthetic promoter; and
a polyadenylation
signal.
|
| CA 2257719 AA |
|
Claim 1
A synthetic DNA plant promoter sequence, said
sequence comprising:
a TATA motif;
a transcription start site; and
a region between said
TATA motif and said start site that is at least 64% GC rich.
|
|
Claim 4
An expression cassette comprising:
a synthetic promoter comprising:
a TATA motif;
a transcription
start site and
a region there between that is at least 64% GC rich;
a
structural gene operatively linked to said promoter; and
a transcription
end site polyadenylation signal.
|
|
Claim 19
A synthetic upstream element having a sequence
of SEQ ID NO:2.
|
|
Claim 20
An expression cassette comprising:
a promoter sequence;
a structural gene operatively linked to said
promoter sequence;
a polyadenylation signal; and
a synthetic upstream
element comprising SEQ ID NO:2 operatively linked to said promoter so that
expression is enhanced.
|
|
Claim 25
A synthetic DNA promoter sequence functional
in plant cells,
wherein said DNA sequence is synthetically created and
non-naturally occurring, consisting of:
a TATA motif;
a transcriptional start site; and
a region between
said TATA motif and said start site that is at least 64% GC rich.
|
|
Claim 26
A synthetic DNA plant promoter sequence
functional in plant cells, said sequence comprising:
a TATA motif;
a transcription start site;
a region between said
TATA motif and said start site that is at least 64% GC rich;
wherein said
promoter sequence is less than 1900 bp.
|
|
US 2001/0047092 A1 and
WO 01/53476 A2
|
|
Claim 1
A plant promoter comprising at least one
synthetic multimeric promoter element region having a nucleotide sequence
selected from the group consisting of:
a) a nucleotide sequence comprising six DRE 1 (SEQ ID NO.: 59), two ABRE1
(SEQ ID NO.: 2), three As-1 (SEQ ID NO.: 7), one GT-2 (SEQ ID NO.: 24), and two
PCNA IIA (SEQ ID NO.: 45) promoter elements;
b) a nucleotide sequence
comprising three DRE 1 (SEQ ID NO.: 59), three ABRE1 (SEQ ID NO.: 2), one As-1
(SEQ ID NO.: 7), two GT-2 (SEQ ID NO.: 24), and two PCNA IIA (SEQ ID NO.: 45)
promoter elements;
c) a nucleotide sequence comprising five DRE 1 (SEQ ID
NO.: 59), three ABRE1 (SEQ ID NO.: 2), two As-1 (SEQ ID NO.: 7), and five GT-2
(SEQ ID NO.: 24) promoter elements;
d) a nucleotide sequence comprising
four DRE 1 (SEQ ID NO.: 59), three ABRE1 (SEQ ID NO.: 2), three GT-2 (SEQ ID
NO.: 24), and one PCNA IIA (SEQ ID NO.: 45) promoter elements;
e) a
nucleotide sequence comprising two DRE 1 (SEQ ID NO.: 59), one ABRE1 (SEQ ID
NO.: 2), five As-1 (SEQ ID NO.: 7), one GT-2 (SEQ ID NO.: 24), and three PCNA
IIA (SEQ ID NO.: 45) promoter elements;
f) a nucleotide sequence comprising
five DRE 1 (SEQ ID NO.: 59), two ABRE1 (SEQ ID NO.: 2), one As-1 (SEQ ID NO.:
7), one GT-2 (SEQ ID NO.: 24), and two PCNA IIA (SEQ ID NO.: 45) promoter
elements;
g) a nucleotide sequence comprising one DRE 1 (SEQ ID NO.: 59),
two ABRE1 (SEQ ID NO.: 2), two As-1 (SEQ ID NO.: 7), and one GT-2 (SEQ ID NO.:
24) promoter elements;
h) a nucleotide sequence comprising two DRE 1, one
ABRE 1 (SEQ ID NO.: 2), three As-1 (SEQ ID NO.: 7), and one GT-2 (SEQ ID NO.:
24) promoter elements; and
i) a nucleotide sequence that hybridizes under
stringent conditions to any of the nucleotide sequences of a), b), c), d), e),
f), g), and h).
|
|
Claim 7
A plant, or its parts, having stably
incorporated into its genome a DNA construct comprising a plant promoter
operably linked to a coding sequence,
said plant promoter comprising at
least one synthetic multimeric promoter element region (SMPER) that enhances
expression of said coding sequence.
|
|
Claim 8
A plant, or its parts, having stably
incorporated into its genome a DNA construct comprising a plant promoter
operably linked to a coding sequences,
said plant promoter comprising at
least one synthetic multimeric promoter element region having a nucleotide
sequence selected from the group consisting of:
a) a nucleotide sequence comprising promoter elements DRE1, ABRE1, DRE1,
As-1, ABRE1, DRE1, GT-2, As-1, DRE1, PCNA IIA, PCNA IIA, DRE1, As-1, and DRE1
sequentially (SEQ ID NO.: 66);
b) a nucleotide sequence comprising
promoter elements DRE1, DRE1, As-1, PCNA IIA, ABRE1, PCNA IIA, ABRE1, DRE1,
GT-2, GT-2, and ABRE1 sequentially (SEQ ID NO.: 67);
c) a nucleotide
sequence comprising promoter elements GT-2, ABRE1, ABRE1, GT-2, As-1, GT-2,
GT-2, DRE1, GT-2, DRE1, DRE1, As-1, DRE1, DRE1, and ABRE1 sequentially (SEQ ID
NO.: 65);
d) a nucleotide sequence comprising promoter elements ABRE1,
ABRE1, GT-2, GT-2, GT-2, DRE1, DRE1, DRE1, DRE1, ABRE1, and PCNA IIA
sequentially (SEQ ID NO.: 68);
e) a nucleotide sequence comprising
promoter elements PCNA IIA, As-1, GT-2, As-1, DRE1, As-1, As-1, PCNA IIA, As-1,
PCNA IIA, DRE1, and ABRE1 sequentially (SEQ ID NO.: 69);
f) a nucleotide
sequence comprising promoter elements As-1, GT-2, DRE1, DRE1, ABRE1, PCNA IIA,
DRE1, PCNA IIA, ABRE1, DRE1, and DRE1 sequentially (SEQ ID NO.: 71);
g) a
nucleotide sequence comprising promoter elements As-1, ABRE 1, GT-2, As-1,
ABRE1, and DRE1 sequentially (SEQ ID NO.: 72);
h) a nucleotide sequence
comprising promoter elements DRE 1, ABRE1, GT-2, DRE1, As-1, As-1, and As-1
sequentially (SEQ ID NO.: 70);
i) a nucleotide sequence set forth in FIGS.
7, 8, 9, 10, 11, 12, 13, or 14 (SEQ ID NOS.: 65-72);
j) a nucleotide
sequence that comprises a variant of a nucleotide sequence set forth in FIGS. 7,
8, 9, 10, 11, 12, 13, or 14 (SEQ ID NOS.: 65-72); and
k) a nucleotide
sequence that hybridizes under stringent conditions to a nucleotide sequence of
(a), (b), (c), (d), (e), (f), (g), (h), (i), or (j).
|
|
Claim 12
A plant cell having stably incorporated into
its genome a DNA construct comprising a plant promoter operably linked to a
coding sequence, said plant promoter comprising at least one synthetic
multimeric promoter element region having a nucleotide sequence selected from
the group consisting of:
a) a nucleotide sequence comprising promoter elements DRE1, ABRE1, DRE1,
As-1, ABRE1, DRE1, GT-2, As-1, DRE1, PCNA IIA, PCNA IIA, DRE1, As-1, and DRE1
sequentially (SEQ ID NO.: 66);
b) a nucleotide sequence comprising
promoter elements DRE1, DRE1, As-1, PCNA IIA, ABRE1, PCNA IIA, ABRE1, DRE1,
GT-2, GT-2, and ABRE1 sequentially (SEQ ID NO.: 67);
c) a nucleotide
sequence comprising promoter elements GT-2, ABRE1, ABRE1, GT-2, As-1, GT-2,
GT-2, DRE1, GT-2, DRE1, DRE1, As-1, DRE1, DRE1, and ABRE1 sequentially (SEQ ID
NO.: 65);
d) a nucleotide sequence comprising promoter elements ABRE 1,
ABRE1, GT-2, GT- 2, GT-2, DRE1, DRE1, DRE1, DRE1, ABRE1 and PCNA IIA
sequentially (SEQ ID NO.: 68);
e) a nucleotide sequence comprising
promoter elements PCNA IIA, As-1, GT-2, As-1, DRE1, As-1, As-1, PCNA IIA, As-1,
PCNA IIA, DRE1, and ABRE1 sequentially (SEQ ID NO.: 69);
f) a nucleotide
sequence comprising promoter elements As-1, GT-2, DRE1, DRE1, ABRE1, PCNA IIA,
DRE1, PCNA IIA, ABRE1, DRE1, and DRE1 sequentially (SEQ ID NO.: 71);
g) a
nucleotide sequence comprising promoter elements As-1, ABRE1, GT-2, As-1, ABRE1,
and DRE1 sequentially (SEQ ID NO.: 72);
h) a nucleotide sequence
comprising promoter elements DRE1, ABRE1, GT-2, DRE1, As-1, As-1, and As-1
sequentially (SEQ ID NO.: 70);
i) a nucleotide sequence set forth in FIGS.
7, 8, 9, 10, 11, 12, 13, or 14 (SEQ ID NOS.: 65-72);
j) a nucleotide
sequence that comprises a variant of a nucleotide sequence set forth in FIGS. 7,
8, 9, 10, 11, 12, 13, or 14 (SEQ ID NOS.: 65-72); and
k) a nucleotide
sequence that hybridizes under stringent conditions to the nucleotide sequence
of (a), (b), (c), (d), (e), (f), (g), (h), (i), or (j).
|
|
Claim 17
A method of selecting promoter elements active
in a tissue of interest, comprising:
a) isolating or synthesizing oligonucleotides representing known or putative
promoter elements or transcription factor binding sites;
b) labeling said
oligonucleotides;
c) pooling said oligonucleotides to create an array which
facilitates screening;
d) hybridizing said oligonucleotides with nuclear
extracts of said tissue of interest; and
e) selecting those
oligonucleotides exhibiting preferential binding to said nuclear extracts.
|
|
Claim 18
A method of creating synthetic multimeric
promoter element regions active in a tissue of interest, comprising:
a) selecting known or putative promoter elements or transcription factor
binding sites which exhibit preferential binding to nuclear extract prepared
from said tissue of interest;
b) combining said selected oligonucletides
in novel arrangements encompassing variation in number of copies, sequential
order, orientation, and spacer regions; and
c) testing said novel
arrangements for their effect on transcription and selecting those demonstrating
enhancement or suppression of linked gene expression.
|
Appendix 3
Claims
Patent granted to Syngenta
Actual granted independent claims
|
EP
637339 B1
|
|
Claim 1
A chemically-inducible plant gene expression
cassette comprising:
a first promoter operatively linked to the alcR regulator sequence
obtainable from Aspergillus nidulans which encodes the AlcR regulator
protein, and
an inducible promoter operatively linked to a target
gene,
the inducible promoter being activated by the regulator protein in
the presence of an effective exogenous inducer whereby application of the
inducer causes expression of the target gene.
|
|
Claim 11
A method for controlling plant gene expression
comprising:
transforming a plant cell with a chemically-inducible plant gene expression
cassette which has
a first promoter operatively linked to the
alcR regulator sequence obtainable from Aspergillus nidulans
which encodes the AlcR regulator protein, and
an inducible promoter
operatively linked to a target gene,
the inducible promoter being
activated by the regulator protein in the presence of an effective exogenous
inducer whereby application of the inducer causes expression of the target gene.
|
|
Claim 12
A chimeric promoter comprising an upstream
region containing
a promoter regulatory sequence obtainable from the alcA gene
promoter of Aspergillus nidulans and
a downstream region
containing a transcription initiation sequence, characterized in that
said upstream and downstream regions are heterologous,
the promoter is
chemically-inducible and
the transcription initiation sequence is
obtainable from the core promoter region of a promoter which is active in plant
cells.
|
Patent granted to Yale University
Actual granted independent claims
|
US
5851796
|
|
Claim 1
A polynucleotide comprising a nucleotide
sequence encoding a tetracycline transactivator fusion protein,
said
protein comprising a prokaryotic tet repressor and a eucaryotic transcriptional
activator protein, and
said polynucleotide molecule being operably linked
to an inducible minimal promoter,
which promoter contains at least one tet
operator sequence.
|
|
Claim 23
A method to inhibit expression of a
heterologous protein in a eucaryotic cell comprising:
a) obtaining a eucaryotic cell comprising:
- a first polynucleotide molecule encoding a tetracycline transactivator
fusion protein,
said protein comprising a procaryotic tet repressor and a
eucaryotic transcriptional activator protein, and
said polynucleotide
molecule being operably linked to an inducible minimal promoter, which promoter
contains at least one tet operator sequence;
- a second polynucleotide molecule encoding the heterologous protein,
said second polynucleotide molecule being operably linked to an inducible
minimal promoter, and
said promoter containing at least one tet operator
sequence; and
b) cultivating the eucaryotic cell in a medium comprising tetracycline or a
tetracycline analogue such that expression of the heterologous protein is
inhibited.
|
|
Claim 25
A method to enhance the expression of a
heterologous protein in a eucaryotic cell comprising:
a) obtaining a eucaryotic cell comprising:
- a first polynucleotide molecule encoding a tetracycline transactivator
fusion protein,
said protein comprising a prokaryotic tet repressor and a
eucaryotic transcriptional activator protein, and
said polynucleotide
molecule being operably linked to an inducible minimal promoter, which promoter
contains at least one tet operator sequence;
- a second polynucleotide molecule encoding the heterologous protein,
said second polynucleotide molecule being operably linked to an inducible
minimal promoter, and
said promoter containing at least one tet operator
sequence; and
b) cultivating the eucaryotic cell in a medium lacking tetracycline or a
tetracycline analogue such that expression of the heterologous protein is
enhanced.
|
|
Claim 26
A method to activate the expression of a
heterologous protein in a eucaryotic cell comprising:
a) obtaining a eucaryotic cell comprising:
- a first polynucleotide molecule encoding a tetracycline transactivator
fusion protein,
said protein comprising a prokaryotic tet repressor and a
eucaryotic transcriptional activator protein, and
said polynucleotide
molecule being operably linked to an inducible minimal promoter, which promoter
contains at least one tet operator sequence;
- a second polynucleotide molecule encoding the heterologous protein,
said second polynucleotide molecule being operably linked to an inducible
minimal promoter, and
said promoter containing at least one tet operator
sequence; and
b) cultivating the eucaryotic cell in a medium lacking tetracycline or a
tetracycline analogue such that expression of the heterologous protein is
activated.
|
|
Claim 27
A kit comprising a carrier means having in
close confinement therein at least two container means, wherein
a) a first container means contains a first polynucleotide molecule encoding
a tetracycline transactivator fusion protein,
said protein comprising a
procaryotic tet repressor and a eucaryotic transcriptional activator protein,
and
said polynucleotide molecule being operably linked to an inducible
minimal promoter, which promoter contains at least one tet operator sequence;
and
b) a second container means contains a second polynucleotide molecule
encoding said inducible minimal promoter,
which promoter contains at least
one tet operator sequence,
which tet operator sequence is strategically
positioned for being operably linked to a heterologous polynucleotide sequence
encoding a polypeptide.
|
|
Claim 28
A kit comprising a carrier means having in
close confinement therein at least two container means, wherein
a) a first container means contains a eucaryotic cell transfected with a
first polynucleotide molecule encoding a tetracycline transactivator fusion
protein,
said protein comprising a procaryotic tet repressor and a
eucaryotic transcriptional activator protein, and
said polynucleotide
molecule being operably linked to an inducible minimal promoter, which promoter
contains at least one tet operator sequence; and
b) a second container
means contains a second polynucleotide molecule comprising an inducible minimal
promoter,
which promoter contains at least one tet operator sequence,
which tet operator sequence is strategically positioned for being operably
linked to a heterologous polynucleotide sequence encoding a heterologous pol
ypeptide.
|
| EP 832254 A1 |
|
Claim 1
A composition of matter comprising a
polynucleotide molecule encoding a tetracycline transactivator fusion
protein,
said protein comprising a prokaryotic tet repressor and a
eucaryotic transcriptional activator protein, and
said polynucleotide
molecule being operably linked to an inducible minimal promoter, which promoter
contains at least one tet operator sequence.
|
|
Claim 11
A method to decrease or shut off expression of
a heterologous protein comprising:
a) transforming a eucaryotic cell with
- a first polynucleotide molecule encoding a tetracycline transactivator
fusion protein,
said protein comprising a prokaryotic tet repressor and a
eucaryotic transcriptional activator protein, and
said polynucleotide
molecule being operably linked to an inducible minimal promoter, which promoter
contains at least one tet operator sequence;
- a second polynucleotide molecule encoding the heterologous protein,
said protein being operably linked to an inducible minimal promoter, and
said promoter containing at least one tet operator sequence; and
b) cultivating the eucaryotic cell in a medium comprising tetracycline or a
tetracycline analogue.
|
|
Claim 13
A method to activate or enhance the expression
of a heterologous protein comprising:
a) transforming a eucaryotic cell with
i. a first polynucleotide molecule encoding tetracycline transactivator
fusion protein,
said protein comprising a prokaryotic tet repressor and a
eucaryotic transcriptional activator protein, and
said polynucleotide
molecule being operably linked to an inducible promoter, which promoter contains
at least one tet operator sequence;
ii. a second polynucleotide molecule encoding the heterologous protein,
said protein being operably linked to an inducible minimal promoter,
and
said promoter containing at least one tet operator sequence; and
b) cultivating the eucaryotic cell in a medium lacking tetracycline or a
tetracycline analogue.
|
|
Claim 17
A composition of matter consisting essentially
of the plasmid pTet-Splice.
|
|
Claim 18
A composition of matter consisting essentially
of the plasmid pTet-tTAK.
|
|
Claim 19
A kit comprising a carrier means having in
close confinement therein at least two container means, wherein
a) a first container means contains a first polynucleotide molecule encoding
a tetracycline transactivator fusion protein,
said protein comprising a
procaryotic tet repressor and a eucaryotic transcriptional activator protein,
and
said polynucleotide molecule being operably linked to an inducible
minimal promoter, which promoter contains at least one tet operator sequence;
and
b) a second container means contains a second polynucleotide molecule
encoding said inducible minimal promoter,
which promoter contains at least
one tet operator sequence, which tet operator sequence is strategically
positioned for being operably linked to a heterologous polynucleotide sequence
encoding a polypeptide.
|
|
Claim 20
A kit comprising a carrier means having in
close confinement therein at least two container means, wherein
a) a first container means contains a eucaryotic cell transfected with a
first polynucleotide molecule encoding a tetracycline transactivator fusion
protein,
said protein comprising a procaryotic tet repressor and a
eucaryotic transcriptional activator protein, and
said polynucleotide
molecule being operably linked to an inducible minimal promoter, which promoter
contains at least one tet operator sequence; and
b) a second container
means contains a second polynucleotide molecule comprising an inducible minimal
promoter,
which promoter contains at least one tet operator sequence, which
tet operator sequence is strategically positioned for being operably linked to a
heterologous polynucleotide sequence encoding a heterologous polypeptide.
|
Patents granted to BASF
Title: Tight control of gene expression in eucaryotic cells
by tetracycline-responsive promoters
Actual granted independent claims
|
US
5464758
|
|
Claim 1
A polynucleotide molecule coding for a
transactivator fusion protein comprising:
a) a prokaryotic Tet repressor and
b) a eucaryotic transcriptional
activator protein domain.
|
|
Claim 10
A polynucleotide molecule coding for a
protein, wherein said polynucleotide is operably linked to a minimal promoter
and at least one tet operator sequence.
|
|
Claim 16
A eucaryotic cell transfected with
a) a first polynucleotide molecule coding for a transactivator fusion pr
otein comprising:
(i) a prokaryotic Tet repressor and
(ii) a
eucaryotic transcriptional activator protein domain; and
b) a second
polynucleotide molecule coding for a protein, wherein said second polynucleotide
molecule is operably linked to a minimal promoter and at least one tet
operator sequence.
|
|
Claim 18
A kit comprising a carrier means having in
close confinement therein at least two container means, wherein
a) a first container means contains a first polynucleotide molecule coding
for a transactivator fusion protein comprising:
(i) a prokaryotic Tet
repressor and
(ii) a eucaryotic transcriptional activator protein domain,
and
b) a second container means contains a second polynucleotide molecule
comprising a minimal promoter operably linked to at least one tet
operator sequence, wherein said minimal promoter is capable of being ligated to
a heterologous gene sequence coding for a polypeptide.
|
|
Claim 19
A kit comprising a carrier means having in
close confinement therein at least two container means, wherein
a) a first container means contains a eucaryotic cell transfected with a
first polynucleotide molecule coding for a transactivator fusion protein
comprising:
(i) a prokaryotic Tet repressor and
(ii) a eucaryotic
transcriptional activator protein domain, and
b) a second container means
contains a second polynucleotide molecule comprising a minimal promoter operably
linked to at least one tet operator sequence, wherein said minimal
promoter is capable of being ligated to a heterologous gene sequence coding for
a polypeptide.
|
|
US
5650298
|
|
Claim 1
An isolated DNA molecule for integrating a
polynucleotide sequence encoding a tetracycline-controllable transactivator
(tTA) at a predetermined location in a second target DNA molecule,
the tTA
comprising a prokaryotic Tet repressor operably linked to a polypeptide which
directly or indirectly activates transcription in eucaryotic cells,
the DNA
molecule comprising a polynucleotide sequence encoding the tTA flanked at 5' and
3' ends by additional polynucleotide sequences of sufficient length for
homologous recombination between the DNA molecule and the second target DNA
molecule at a predetermined location.
|
|
Claim 11
An isolated DNA molecule for integrating a
polynucleotide sequence encoding a tetracycline-controllable transactivator
(tTA) and a tTA-responsive promoter within a predetermined gene of interest in a
second target DNA molecule, the DNA molecule comprising:
a) a first polynucleotide sequence comprising a 5' flanking regulatory region
of the gene of interest, operably linked to;
b) a second polynucleotide
sequence encoding a tTA,
the tTA comprising a prokaryotic Tet repressor
operably linked to a polypeptide which directly or indirectly activates
transcription in eucaryotic cells; and
c) a third polynucleotide sequence
comprising a tTA-responsive promoter, operably linked to;
d) a fourth
polynucleotide sequence comprising at least a portion of a coding region of the
gene of interest;
wherein the first and fourth polynucleotide sequences are
of sufficient length for homologous recombination between the DNA molecule and
the gene of interest in the second target DNA molecule such that expression of
the tTA is controlled by 5' regulatory elements of the gene of interest and
expression of the gene of interest is controlled by the tTA-responsive promoter.
|
| CA 2165162 |
|
Claim 1
Worded the same as claim 1 of
US 5 650 298.
|
|
Claim 11
Worded the same as claim 11
of US 5 650 298.
|
|
Claim 42
Use of tetracycline or a tetracycline analogue
for the inhibition of a second transgene in a transgenic animal, said animal
having:
a) a first transgene comprising a polynucleotide sequence encoding a
tetracycline-controllable transactivator (tTA),
the tTA comprising a
prokaryotic Tet repressor operably linked to a polypeptide which directly or
indirectly activates transcription in eucaryotic cells; and
b) the second
transgene comprising a gene of interest operably linked to a tTA-responsive
promoter.
|
|
Claim 43
The use of tetracycline or a tetracycline
analogue for the inhibition of transcription of a second transgene in a
transgenic animal, said animal having:
a) a polynucleotide sequence encoding a tetracycline-controllable
transactivator (tTA),
the tTA comprising a prokaryotic Tet repressor
operably linked to a polypeptide which directly or indirectly activates
transcription in eucaryotic cells, wherein the first transgene is integrated by
homologous recombination at a predetermined location within a chromosome within
cells of the animal; and
b) the second transgene comprising a gene of
interest operably linked to a tTA-responsive promoter.
|
|
Claim 44
Use of tetracycline or a tetracycline analogue
for inhibiting transcription of the gene of interest in a transgenic animal
having a transgene comprising a polynucleotide sequence encoding a
tetracycline-controllable transactivator (tTA) and a tTA-responsive
promoter,
wherein the transgene is integrated by homologous recombination
at a predetermined location within a gene of interest within cells of the animal
such that expression of the tTA is controlled by 5' regulatory elements of the
gene of interest and expression of the gene of interest is controlled by the
tTA-responsive promoter.
|
|
Claim 48
A method for producing a non-human transgenic
animal comprising:
a) introducing a DNA molecule encoding the tTA into a fertilized oocyte;
b) implanting the fertilized oocyte in a pseudopregnant foster mother;
and
c) allowing the fertilized oocyte to develop into the non-human
transgenic animal to thereby produce the non-human transgenic animal.
|
| AU 684524 B2 |
|
Claim 1
Worded the same as Claim 1 of
Canadian patent and US 5 650 298.
|
|
Claim 11
Worded the same as Claim 11
of Canadian pat ent and US 5 650 298.
|
|
Claim 42
A non-human transgenic animal having a
transgene comprising
a polynucleotide sequence encoding a
tetracycline-controllable transactivator (tTA),
the tTA comprising a
prokaryotic Tet repressor operably linked to a polypeptide which directly or
indirectly activates transcription in eucaryotic cells.
|
|
Claim 52
A non-human transgenic animal having a
transgene comprising
a polynucleotide sequence encoding a
tetracycline-controllable transactivator (tTA),
the tTA comprising a
prokaryotic Tet repressor operably linked to a polypeptide which directly or
indirectly activates transcription in eucaryotic cells,
wherein the
transgene is integrated by homologous recombination at a predetermined location
within a chromosome within cells of the animal.
|
|
Claim 55
A transgenic animal having a transgene
comprising
a polynucleotide sequence encoding a tetracycline-controllable
transactivator (tTA) and a tTA-responsive promoter,
wherein the transgene
is integrated by homologous recombination at a predetermined location within a
gene of interest within cells of the animal such that expression of the tTA is
controlled by 5' regulatory elements of the gene of interest and expression of
the gene of interest is controlled by the tTA-responsive promoter.
|
|
US
5589362
|
|
Claim 1
An isolated nucleic acid molecule encoding a
fusion protein which regulates transcription,
the fusion protein comprising
a Tet repressor having at least one amino acid mutation that confers on the
fusion protein an ability to bind a class B tet operator sequence
having a nucleotide substitution at position +4 or +6, operatively linked to a
polypeptide which regulates transcription in eukaryotic cells.
|
|
Claim 13
A method for regulating transcription of a
tet operator-linked gene in an isolated cell, comprising:
a) introducing into the isolated cell a nucleic acid molecule encoding a
fusion protein which regulates transcription,
the fusion protein comprising
a Tet repressor having at least one amino acid mutation that confers on the
fusion protein an ability to bind a class B tet operator sequence having a
nucleotide substitution at position 14 or 16, operatively linked to a
polypeptide which regulates transcription in eukaryotic cells; and
b)
modulating the concentration of a tetracycline, or analogue thereof, in contact
with the isolated cell.
|
|
US
5654168
|
|
Claim 1
An isolated nucleic acid encoding a fusion
protein which activates transcription, the fusion protein
comprising
a first polypeptide which binds to a tet operator
sequence in the presence of tetracycline or a tetracycline analogue operatively
linked
to a second polypeptide which activates transcription in eukaryotic
cells.
|
|
Claim 26
A kit comprising a carrier means having in
close confinement therein at least two container means comprising:
a) a first container means containing a first nucleic acid encoding a fusion
protein which activates transcription, the fusion protein
comprising
a polypeptide which binds to a first class of tet
operator sequence in the presence of tetracycline or a tetracycline analogue
operatively linked to a polypeptide which activates transcription in eukaryotic
cells; and
b) a second container means containing a second nucleic acid
comprising a first cloning site for introduction of a first nucleotide sequence
to be transcribed operatively linked to at least one tet operator
sequence of a first class type.
|
|
US
5789156
|
|
Claim 1
An isolated nucleic acid encoding a fusion
protein which inhibits transcription in eukaryotic cells, the
fusion protein comprising a first polypeptide which binds to tet operator
sequences, operatively linked to a heterologous second polypeptide which
inhibits transcription in eukaryotic cells.
|
|
Claim 46
A kit comprising a carrier means having in
close confinement therein at least two container means comprising:
a) a first container means containing a first nucleic acid encoding a fusion
protein which inhibits transcription in eukaryotic cells, the
fusion protein comprising
a first polypeptide which binds to tet
operator sequences either
i. in the presence but not the absence of tetracycline or a tetracycline
analogue or
ii. in the absence but not the presence of tetracycline or a tetracycline
analogue,
operatively linked to a heterologous second polypeptide which
inhibits transcription in eukaryotic cells, or
a eukaryotic cell line into
which said first nucleic acid has been stably introduced; and
b) a second container means containing a second nucleic acid comprising a
cloning site for introduction of a nucleotide sequence to be transcribed
operatively linked to at least one tet operator sequence.
|
Patents entitled: "Methods for regulating gene expression"
|
US
5814618
|
|
Claim 1
A method for regulating expression of a
tet operator-linked gene in a cell of a subject, comprising:
a) introducing into the cell a nucleic acid molecule encoding a fusion
protein which inhibits transcription in eukaryotic cells, the
fusion protein comprising
a first polypeptide which binds to a
tet operator sequence, operatively linked to a heterologous second
polypeptide which inhibits transcription in eukaryotic cells; and
b)
modulating the concentration of a tetracycline, or analogue thereof, in the
subject.
|
|
Claim 17
A method for regulating expression of a gene
in a cell of a subject, comprising:
a) obtaining the cell from the subject;
b) introducing into the cell a
first nucleic acid molecule which operatively links a gene to at least one
tet operator sequence;
c) introducing into the cell a second
nucleic acid molecule encoding a fusion protein which inhibits
transcription, the fusion protein comprising
a first polypeptide which
binds to a tet operator sequence, operatively linked to a second
polypeptide which inhibits transcription in eukaryotic cells, to form a modified
cell;
d) administering the modified cell to the subject; and
e)
modulating the concentration of a tetracycline, or analogue thereof, in the
subject.
|
|
US
5888981
|
|
Claim 1
A method for regulating expression of a
tet operator-linked gene in a cell of a subject, comprising:
a) introducing into the cell a nucleic acid molecule encoding a
tetracycline-controllable transactivator (tTA), the tTA comprising
a Tet
repressor operably linked to a polypeptide which directly or indirectly
activates transcription in eucaryotic cells; and
b) modulating the
concentration of a tetracycline, or analogue thereof, in the subject.
|
|
Claim 10
A method for regulating expression of a gene
in a cell of a subject, comprising:
a) obtaining the cell from the subject;
b) introducing into the cell a
first nucleic acid molecule which operatively links a gene to at least one
tet operator sequence;
c) introducing into the cell a second
nucleic acid molecule encoding a te tracycline-controllable transactivator
(tTA), the tTA comprising
a Tet repressor operably linked to a polypeptide
which directly or indirectly activates transcription in eucaryotic cells, to
form a modified cell;
d) administering the modified cell to the subject;
and
e) modulating the concentration of a tetracycline, or analogue
thereof, in the subject.
|
|
US 6004941
|
|
Claim 1
A method for regulating expression of a tet
operator-linked gene in a cell of a subject, comprising:
a) introducing into the cell a nucleic acid molecule encoding a fusion
protein which activates transcription, the fusion protein comprising
a
first polypeptide which binds to a tet operator sequence in the
presence of tetracycline or a tetracycline analogue operatively linked to
a second polypeptide which activates transcription in eukaryotic cells; and
b) modulating the concentration of a tetracycline, or analogue thereof, in
the subject, such that expression of a tet operator-linked gene in a
cell of the subject is regulated.
|
|
Claim 16
A method for regulating expression of a gene
in a cell of a subject, comprising:
a) obtaining the cell from the subject;
b) introducing into the cell a
first nucleic acid molecule which operatively links a gene to at least one tet
operator sequence;
c) introducing into the cell a second nucleic acid
molecule encoding a fusion protein which activates transcription, the fusion
protein comprising
a first polypeptide which binds to a tet
operator sequence in the presence of tetracycline or a tetracycline analogue
operatively linked to
a second polypeptide which activates transcription
in eukaryotic cells, to form a modified cell;
d) administering the modified
cell to the subject; and
e) modulating the concentration of a
tetracycline, or analogue thereof, in the subject such that expression of the
gene which is operatively linked to at least one tet operator sequence
is regulated in a cell of the subject.
|
|
Claim 28
An isolated recombinant vector for coordinate,
bidirectional transcription of a first and a second nucleotide sequence,
the vector comprising a nucleotide sequence comprising in a 5' to 3' direction:
a) a first cloning site for introduction of a first nucleotide sequence to be
transcribed, operatively linked to
b) at least one tet operator
sequence, operatively linked to
c) a second cloning site for introduction
of a second nucleotide sequence transcribed,
the vector further comprising
additional regulatory sequences such that the vector is sufficient for use in
eukaryotic cells,
wherein transcription of a first and second nucleotide
sequence introduced into the vector proceeds in opposite directions relative to
the at least one tet operator sequence.
|
|
Claim 35
An isolated nucleic acid composition
comprising at least one recombinant vector for independent regulation of
transcription of a first and a second nucleotide sequence, the nucleic acid
composition comprising nucleotide sequences comprising:
a) a first cloning site for introduction of a first nucleotide sequence to be
transcribed, operatively linked to at least one tet operator sequence
of a first class type; and
b) a second cloning site for introduction of a
second nucleotide sequence to be transcribed, operatively linked to at least one
tet operator sequence of a second class type.
|
Transgenic mice with a tetracycline-responsive transcriptional regulator
Patents granted to BASF
|
US
5859310
|
|
Claim 1
A transgenic mouse having a transgene
integrated into the genome of the mouse and also having a tet operator-linked
gene in the genome of the mouse, wherein:
a)the transgene comprises a transcriptional regulatory element functional in
cells of the mouse operatively linked to a polynucleotide sequence encoding a
tetracycline-controllable transactivator fusion protein (tTA),
b) said
fusion protein comprises a Tet repressor operably linked to a polypeptide which
directly or indirectly activates transcription of said tet
operator-linked gene in eucaryotic cells,
c) said tet
operator-linked gene confers a detectable and functional phenotype on the
mouse when expressed in cells of the mouse,
d) said transgene is expressed
in cells of the mouse at a level sufficient to produce amounts of said fusion
protein that are sufficient to activate transcription of the tet
operator-linked gene; and
e) in the absence of tetracycline or a
tetracycline analogue in the mouse, said fusion protein binds to the
tet operator-linked gene and activates transcription of the tet
operator linked gene such that the tet operator-linked gene is
expressed at a level sufficient to confer the detectable and functional
phenotype on the mouse,
wherein the level of expression of tet operator-linked gene can be
down modulated by administering tetracycline or a tetracycline analogue to the
mouse.
|
|
Claim 13
A transgenic mouse having a transgene
integrated into the genome of the mouse, wherein:
a) the transgene comprises a polynucleotide sequence encoding a tet
racycline-controllable transactivator fusion protein (tTA) and a tTA-responsive
promoter,
b) said fusion protein comprises a Tet repressor operably linked
to a polypeptide which directly or indirectly activates transcription of a gene
of interest in eucaryotic cells,
c) the transgene is integrated by
homologous recombination at a predetermined location within a said gene of
interest within cells of the mouse such that expression of the fusion protein is
controlled by 5' regulatory elements of the gene of interest and expression of
the gene of interest is controlled by the tTA-responsive promoter, expression of
the gene of interest confers a detectable and functional phenotype on the mouse,
d) said transgene is expressed in cells of the mouse at a level sufficient
to produce amounts of said fusion protein that are sufficient to activate
transcription of the gene of interest linked to the tTA-responsive promoter, and
e) in the absence of tetracycline or a tetracycline analogue in the mouse,
said fusion protein binds to the tTA-responsive promoter and activates
transcription of the gene of interest such that the gene of interest is
expressed at a level sufficient to confer the detectable and functional
phenotype on the mouse,
f) wherein the level of expression of the gene of
interest can be down modulated by administering tetracycline or a tetracycline
analogue to the mouse.
|
|
Claim 20
A transgenic mouse having a transgene
integrated into the genome of the mouse, wherein:
a) the transgene comprises a transcriptional regulatory element functional in
cells of the mouse operatively linked to a polynucleotide sequence encoding a
tetracycline-controllable transactivator fusion protein (tTA),
b) said
fusion protein comprises a Tet repressor operably linked to a polypeptide which
directly or indirectly activates transcription of a tet operator-linked gene in
eucaryotic cells, and
c) said fusion protein is expressed in cells of the
mouse.
|
|
US
5866755
|
|
Claim 1
A transgenic mouse having a transgene
integrated into the genome of the mouse and also having a tet
operator-linked gene in the genome of the mouse, wherein:
a) the transgene comprises a transcriptional regulatory element functional in
cells of the mouse operatively linked to a polynucleotide sequence encoding a
fusion protein which inhibits transcription of said tet operator linked
gene,
b) said fusion protein comprises a first polypeptide that is a Tet
repressor operably linked to a heterologous second polypeptide which inhibits
transcription of said tet operator-linked gene in eucaryotic cells,
c) said tet operator-linked gene confers a detectable and functional
phenotype on the mouse when expressed in cells of the mouse,
d) said
transgene is expressed in cells of the mouse at a level sufficient to produce
amounts of said fusion protein that are sufficient to inhibit transcription of
the tet operator-linked gene; and
e) in the absence of
tetracycline or a tetracycline analogue in the mouse, said fusion protein binds
to the tet operator-linked gene and inhibits transcription of the
tet operator linked gene,
f) wherein the level of expression of
the tet operator-linked gene can be upregulated by administering
tetracycline or a tetracycline analogue to the mouse.
|
|
Claim 2
A transgenic mouse having a transgene
integrated into the genome of the mouse and also having a tet
operator-linked gene in the genome of the mouse, wherein:
a) the transgene comprises a transcriptional regulatory element functional in
cells of the mouse operatively linked to a polynucleotide sequence encoding a
fusion protein which inhibits transcription of said tet operator linked
gene,
b) said fusion protein comprises a first polypeptide that is a
mutated Tet repressor that binds to tet operator sequences in the
presence, but not the absence, of tetracycline or a tetracycline analogue,
operably linked to a heterologous second polypeptide which inhibits
transcription of
c) said tet operator-linked gene in eucaryotic
cells,
d) said tet operator-linked gene confers a detectable and
functional phenotype on the mouse when expressed in cells of the mouse,
e)
said transgene is expressed in cells of the mouse at a level sufficient to
produce amounts of said fusion protein that are sufficient to inhibit
transcription of the tet operator-linked gene; and
f) in the
presence of tetracycline or a tetracycline analogue in the mouse, said fusion
protein binds to the tet operator-linked gene and inhibits
transcription of the tet operator linked gene,
g) wherein the
level of expression of the tet operator-linked gene can be upregulated
by depleting tetracycline or a tetracycline analogue from the mouse.
|
|
Claim 3
A transgenic mouse having a transgene
integrated into the genome of the mouse, wherein:
a) the transgene comprises a transcriptional regulatory element functional in
cells of the mouse operatively linked to a polynucleotide sequence encoding a
fusion protein which inhibits transcription of a tet operator-linked
gene,
b) the fusion protein comprising a first polypeptide that is a Tet
repressor or, a mutated Tet repressor that binds to a tet operator
sequence, operatively linked to a second polypeptide which inhibits
transcription in eukaryotic cells, and
c) said fusion protein is expressed
in cells of the mouse.
|
|
US
5912411
|
|
Claim 1
A transgenic mouse having a transgene
integrated into the genome of the mouse and also having a tet
operator-linked gene in the genome of the mouse, wherein:
a) the transgene comprises a transcriptional regulatory element functional in
cells of the mouse operatively linked to a polynucleotide sequence encoding a
fusion protein which activates transcription of said tet
operator-linked gene,
b) the fusion protein comprises a first polypeptide
which is a mutated Tet repressor that binds to a tet operator sequence
in the presence of tetracycline or a tetracycline analogue operatively linked to
a second polypeptide which activates transcription in eukaryotic cells,
c)
said tet operator-linked gene confers a detectable and functional
phenotype on the mouse when expressed in cells of the mouse,
d) said
transgene is expressed in cells of the mouse at a level sufficient to produce
amounts of said fusion protein that are sufficient to activate transcription of
the tet operator-linked gene; and
e) in the presence of
tetracycline or a tetracycline analogue in the mouse, said fusion protein binds
to the tet operator-linked gene and activ ates transcription of the
tet operator-linked gene such that the tet operator-linked
gene is expressed at a level sufficient to confer the detectable and functional
phenotype on the mouse,
f) wherein the level of expression of the
tet operator-linked gene cap be down modulated by depleting
tetracycline or a tetracycline analogue from the mouse.
|
|
Claim 35
A transgenic mouse having a transgene
integrated into the genome of the mouse, wherein:
a) the transgene comprises a transcriptional regulatory element functional in
cells of the mouse operatively linked to a polynucleotide sequence encoding a
fusion protein which activates transcription of a tet operator-linked
gene,
b) the fusion protein comprising a first polypeptide which is a
mutated Tet repressor that binds to a tet operator sequence in the
presence of tetracycline or a tetracycline analogue operatively linked to a
second po lypeptide which activates transcription in eukaryotic cells, and
c) said fusion protein is expressed in cells of the mouse.
|
|
US
5922927
|
|
Claim 1
A method for producing a transgenic mouse,
comprising:
a) introducing into a fertilized oocyte of a mouse a DNA molecule encoding a
tetracycline-controllable transactivator (tTA),
the tTA comprising a
prokaryotic Tet repressor operably linked to a polypeptide which directly or
indirectly activates transcription in eucaryotic cells;
b) implanting the
fertilized oocyte in a pseudopregnant foster mother; and
c) allowing the
fertilized oocyte to develop into a transgenic mouse to thereby produce the
transgenic mouse, wherein said tTA is expressed in cells of the mouse at a level
sufficient to transactivate a tet operator-linked gene.
|
|
Claim 2
A method for producing a transgenic mouse
having a transgene encoding a tetracycline-controllable transactivator (tTA)
integrated at a predetermined location within chromosomal DNA of cells of the
mouse, comprising:
a) introducing into a population of embryonic stem cells of a mouse a DNA
molecule encoding a tTA,
the tTA comprising a prokaryotic Tet repressor
operably linked to a polypeptide which directly or indirectly activates
transcription in eucaryotic cells,
the DNA molecule comprising a
polynucleotide sequence encoding the tTA flanked at 5' and 3' ends by additional
polynucleotide sequences of sufficient length for homologous recombination
between the DNA molecule and a second target DNA molecule at a predetermined
location within chromosomal DNA of cells of the mouse, under conditions suitable
for homologous recombination between the DNA molecule encoding the tTA and
chromosomal DNA within the cell;
b) selecting an embryonic stem cell in
which DNA encoding the tTA has integrated at a predetermined location within the
chromosomal DNA of the cell;
c) implanting the embryonic stem cell into a
blastocyst;
d) implanting the blastocyst into a pseudopregnant foster
mother; and
e) allowing the blastocyst to develop into a transgenic mouse
to thereby produce the transgenic mouse,
wherein said tTA is expressed in
cells of the mouse at a level sufficient to transactivate a tet
operator-linked gene.
|
|
Claim 3
A method for producing a transgenic mouse
having a transgene encoding a tetrac ycline-controllable transactivator (tTA)
and a tTA-responsive promoter integrated at a predetermined location within a
gene of interest in cells of the mouse, comprising:
a) introducing into a population of embryonic stem cells of a mouse a DNA
molecule encoding a tTA, the DNA molecule comprising:
- a first polynucleotide sequence comprising a 5' flanking regulatory region
of the gene of interest, operably linked to;
- a second polynucleotide sequence encoding a tTA, the tTA comprising a
prokaryotic Tet repressor operably linked to a polypeptide which directly or
indirectly activates transcription in eucaryotic cells; and
- a third polynucleotide sequence comprising a tTA-responsive promoter,
operably linked to;
- a fourth polynucleotide sequence comprising at least a portion of a coding
region of the gene of interest;
- wherein the first and fourth polynucleotide sequences are of sufficient
length for homologous recombination between the DNA molecule and the gene of
interest such that expression of the tTA is controlled by 5' regulatory elements
of the gene of interest and expression of the gene of interest is controlled by
the tTA-responsive promoter, under conditions suitable for homologous
recombination between the DNA molecule encoding the tTA and the gene of interest
within the cell;
b) selecting an embryonic stem cell in which DNA encoding the tTA has
integrated at a predetermined location within the gene of interest in the cell;
c) implanting the embryonic stem cell into a blastocyst;
d)
implanting the blastocyst into a pseudopregnant foster mother; and
e)
allowing the blastocyst to develop into a transgenic mouse to thereby produc e
the transgenic mouse,
wherein said gene of interest confers a detectable
and functional phenotype on the mouse when expressed in cells of the transgenic
mouse, said tTA is expressed in cells of the transgenic mouse at a level
sufficient to activate transcription of the gene of interest; and
in the
absence of tetracycline or a tetracycline analogue in the mouse, said tTA binds
to the tTA responsive promoter operably linked to the gene of interest and
activates transcription of the gene of interest such that the gene of interest
is expressed at a level sufficient to confer the detectable and functional
phenotype on the mouse,
wherein the level of expression of the gene of
interest can be down modulated by administering tetracycline or a tetracycline
analogue to the mouse.
|
|
US
6136954
|
|
Claim 1
A fusion protein which
activates transcription comprising:
a first polypeptide
which binds to a tet operator sequence in the presence of tetracycline
or a tetracycline analogue operatively linked
to
a second polypeptide
which activates transcription in eukaryotic cells.
|
|
US
6271348
|
|
Claim 1
A fusion protein which
inhibits transcription in eukaryotic cells, the fusion protein
comprising:
a first polypeptide which binds to tet operator
sequences, operatively linked to
a heterologous second polypeptide which
inhibits transcription in eukaryotic cells.
|
Patents entitled: "Transgenic organisms having tetracycline-regulated
transcriptional regulatory systems"
|
US
6242667
|
|
Claim 1
A transgenic plant having a transgene
integrated into the genome of the plant and also having a tet
operator-linked gene in the genome of the plant, wherein:
a) the transgene comprises:
a transcriptional regulatory element
functional in cells of the plant operatively linked to a polynucleotide sequence
encoding a fusion protein which activates transcription of said tet
operator-linked gene,
the fusion protein comprises
a) a first polypeptide which is a mutated
Tet repressor that binds to a te t operator sequence in the presence of
tetracycline or a tetracycline analogue operatively linked to
b) a second
polypeptide which activates transcription in eukaryotic cells,
b) said tet operator-linked gene confers a detectable and functional
phenotype on the plant when expressed in cells of the plant,
c) said
transgene is expressed in cells of the plant at a level sufficient to produce
amounts of said fusion protein that are sufficient to activate transcription of
the tet operator-linked gene; and
d) in the presence of
tetracycline or a tetracycline analogue in the plant, said fusion protein binds
to the tet operator-linked gene and activates transcription of the
tet operator-linked gene such that the tet operator-linked
gene is expressed at a level sufficient to confer the detectable and functional
phenotype on the plant,
wherein the level of expression of the tet
operator-linked gene can be down modulated by depleting tetracycline or a
tetracycline analogue from the plant.
|
|
Claim 2
A transgenic plant having a transgene
integrated into the genome of the plant, wherein:
a) the transgene comprises
a transcriptional regulatory element
functional in cells of the plant operatively linked to a polynucleotide sequence
encoding a fusion protein which activates transcription of a tet
operator linked gene,
the fusion protein comprising
a) a first polypeptide which is a mutated
Tet repressor that binds to a tet operator sequence in the presence of
tetracycline or a tetracycline analogue operatively linked to
b) a second
polypeptide which activates transcription in eukaryotic cells, and
said
fusion protein is expressed in cells of the plant.
|
|
Claim 3
A transgenic plant having a transgene
integrated into the genome of the plant and also having a tet
operator-linked gene in the genome of the plant, wherein:
the transgene comprises
a transcriptional regulatory element functional
in cells of the plant operatively linked to a polynucleotide sequence encoding a
fusion protein which inhibits transcription of said tet operator linked
gene,
the fusion protein comprises
a first polypeptide which is a
mutated Tet repressor that binds to a tet operator sequence in the
presence of tetracycline or a tetracycline analogue operatively linked to
a second polypeptide which inhibits transcription in eukaryotic cells,
said tet operator-linked gene confers a detectable and functional
phenotype on the plant when expressed in cells of the plant,
said
transgene is expressed in cells of the plant at a level sufficient to produce
amounts of said fusion protein that are sufficient to inhibit transcription of
the tet operator-linked gene; and
in the presence of tetracycline
or a tetracycline analogue in the plant, said fusion protein binds to the
tet operator-linked gene and inhibits transcription of the tet
operator-linked gene, wherein the level of expression of the tet
operator-linked gene can be upregulated by depleting tetracycline or a
tetracycline analogue from the plant.
|
|
Claim 4
A transgenic plant having a transgene
integrated into the genome of the plant, wherein: the transgene comprises a
transcriptional regulatory element functional in cells of the plant operatively
linked to a polynucleotide sequence encoding a fusion protein which inhibits
transcription of a tet operator linked gene, the fusion protein comprising a
first polypeptide which is a mutated Tet repressor that binds to a tet operator
sequence in the presence of tetracycline or a tetracycline analogue operatively
linked to a second polypeptide which inhibits transcription in eukaryotic cells,
and said fusion protein is expressed in cells of the plant.
|
|
Claim 5
A transgenic plant having a transgene
integrated into the genome of the plant and also having a tet operator-linked
gene in the genome of the plant, wherein: the transgene comprises a
transcriptional regulatory element functional in cel ls of the plant operatively
linked to a polynucleotide sequence encoding a fusion protein which inhibits
transcription of said tet operator linked gene, said fusion protein comprises a
first polypeptide that is a Tet repressor, operably linked to a heterologous
second polypeptide which inhibits transcription of said tet operator-linked gene
in eukaryotic cells, said tet operator-linked gene confers a detectable and
functional phenotype on the plant when expressed in cells of the plant, said
transgene is expressed in cells of the plant at a level sufficient to produce
amounts of said fusion protein that are sufficient to inhibit transcription of
the tet operator-linked gene; and in the absence of tetracycline or a
tetracycline analogue in the plant, said fusion protein binds to the tet
operator-linked gene and inhibits transcription of the tet operator linked gene,
wherein the level of expression of the tet operator-linked gene can be
upregulated by administering tetracycline or a tetracycline analogue to the
plant.
|
|
Claim 6
A transgenic plant having a transgene
integrated into the genome of the plant, wherein: the transgene comprises a
transcriptional regulatory element functional in cells of the plant operatively
linked to a polynucleotide sequence encoding a fusion protein which inhibits
transcription of a tet operator linked gene, the fusion protein comprising a
first polypeptide which is a Tet repressor, operatively linked to a second
polypeptide which inhibits transcription in eukaryotic cells, and said fusion
protein is expressed in cells of the plant.
|
|
US
6252136
|
|
Claim 1
A transgenic plant having a transgene
integrated into the genome of the plant and also having a tet operator-linked
gene in the genome of the plant, wherein: the transgene comprises a
transcriptional regulatory element functional in c ells of the plant operatively
linked to a polynucleotide sequence encoding a fusion protein which activates
transcription of said tet operator linked gene, the fusion protein comprises a
first polypeptide which is a Tet repressor operatively linked to a second
polypeptide which directly or indirectly activates transcription in plant cells,
said tet operator-linked gene confers a detectable and functional phenotype on
the plant when expressed in cells of the plant, said transgene is expressed in
cells of the plant at a level sufficient to produce amounts of said fusion
protein that are sufficient to activate transcription of the tet operator-linked
gene; and in the absence of tetracycline or a tetracycline analogue in the
plant, said fusion protein binds to the tet operator-linked gene and activates
transcription of the tet operator linked gene such that the tet operator-linked
gene is expressed at a level sufficient to confer the detectable and functional
phenotype on the plant, wherein the level of expression of the tet
operator-linked gene can be down modulated by administering tetracycline or a
tetracycline analogue to the pl ant.
|
|
Claim 7
A transgenic plant having a transgene
integrated into the genome of he plant, wherein: the transgene comprises a
transcriptional regulatory element fun ctional in cells of the plant operatively
linked to a polynucleotide sequence encoding a fusion protein which activates
transcription of a tet operator linked gene, the fusion protein comprising a
first polypeptide which is a Tet repressor, operatively linked to a second
polypeptide which directly or indirectly activates transcription in plant cells,
and said fusion protein is expressed in cells of the plant.
|
| CA 2193122 |
|
Claim 1
An isolated nucleic acid encoding a fusion
protein which activates transcription,
the fusion protein comprising a
first polypeptide which binds to a tet operator sequence in the
presence, but not the absence, of tetracycline or a tetracycline analogue
operatively linked to a second polypeptide which activates transcription in
eukaryotic cells.
|
|
Claim 12
A fusion protein which activates transcription
comprising a first polypeptide which binds to a tet operator sequence
in the presence, but not the absence, of tetracycline or a tetracycline analogue
operatively linked to a second polypeptide which activates transcription in
eukaryotic cells .
|
|
Claim 35
An isolated nucleic acid encoding a fusion
protein which inhibits transcription in eukaryotic cells,
the fusion
protein comprising a first polypeptide which binds to a tet operator
sequence operatively linked to a heterologous second polypeptide which inhibits
transcription in eukaryotic cells.
|
|
Claim 51
A fusion protein which inhibits transcription
in eukaryotic cells, comprising a first polypeptide which binds to a
tet operator sequence operatively linked to a heterologous second
polypeptide which inhibits transcription in eukaryotic cells.
|
|
Claim 73
A host cell comprising:
a) a first nucleic acid encoding a first fusion protein which activates
transcription,
the first fusion protein comprising a first polypeptide
which binds to a tet operator sequence operatively linked to a second
polypeptide which activates transcription in eukaryotic cells;
b) a second
nucleic acid encoding a second fusion protein which inhibits
transcription,
the second fusion protein comprising a third polypeptide
which binds to a tet operator sequence operatively linked to a fourth
polypeptide which inhibits transcription in eukaryotic cells; and
c) a
third nucleic acid molecule comprising a nucleotide sequence to be transcribed
operatively linked to at least one tet operator sequence.
|
|
Claim 76
A non-human transgenic organism comprising:
a) a first transgene encoding a first fusion protein which activates
transcription,
the first fusion protein comprising a first polypeptide
which binds to a tet operator sequence operatively linked to a second
polypeptide which activates transcription in eukaryotic cells;
b) a second
transgene encoding a second fusion protein which inhibits transcription,
the second fusion protein comprising a third polypeptide which binds to a
tet operator sequence operatively linked to a fourth polypeptide which
inhibits transcription in eukaryotic cells; and
c) a third transgene
comprising a nucleotide sequence to be transcribed operatively linked to at
least one tet operator sequence.
|
|
Claim 79
A recombinant vector for coordinate,
bidirectional transcription of a first and a second nucleotide sequence to be
transcribed,
the vector comprising a nucleotide sequence comprising in a 5'
to 3' direction:
a first cloning site for introduction of a first
nucleotide sequence to be transcribed,
which is operatively linked to at
least one tet operator sequence,
which is operatively linked to a
second cloning site for introduction of a second nucleotide sequence to be
transcribed,
wherein transcription of the first and second nucleotide
sequence introduced into the vector proceeds in opposite directions relative to
the at least one tet operator sequence.
|
|
Claim 83
A composition of matter comprising at least
one recombinant vector for independent regulation of transcription of a first
and a second nucleotide sequence to be transcribed, the at least one vector
comprising a nucleotide sequence comprising:
a) a first cloning site for introduction of a first nucleotide sequence to be
transcribed, operatively linked to at least one tet operator sequence
of a first class type; and
b) a second cloning site for introduction of a
second nucleotide sequence to be transcribed, operatively linked to at least one
tet operator sequence of a second class type.
|
|
Claim 87
A kit comprising a carrier means having in
close confinement therein at least two container means comprising:
a) a first container means containing a first nucleic acid encoding a fusion
protein which activates transcription,
the fusion protein comprising a
first polypeptide which binds to a tet operator sequence in the
presence of tetracycline or a tetracycline analogue operatively linked to a
second polypeptide which activates transcription in eukaryotic cells; and
b) a second container means containing a second nucleic acid comprising a
cloning site for introduction of a nucleotide sequence to be transcribed
operatively linked to at least one tet operator sequence.
|
| EP 1092771 |
|
Claim 1
An isolated nucleic acid encoding a fusion
protein which inhibits transcription in eukaryotic cells,
the fusion
protein comprising:
a) a first polypeptide which is a Tet repressor that binds to a tet
operator sequence in the absence but not the presence of tetracycline or a
tetracycline analogue operatively linked to
b) a heterologous second
polypeptide which inhibits transcription in eukaryotic cells.
|
1. Promoters based on glucocorticoid receptor (GR)
Actual granted independent claims
|
US
5512483
|
|
Claim 1
An expression vector adapted for replication in
an animal cell comprising a glucocorticoid responsive promoter,
said promoter comprising
a) a plurality of at least 5 glucocorticoid response elements (GREs),
b) a viral or mammalian TATA box, and
c) a viral or mammalian initiator
element with a transcriptional initiator site located from 20 to 50 bases from
said TATA box,
said promoter lacking upstream elements which bind nuclear factor I, and
said vector further comprising a restriction endonuclease site downstream
from said promoter for insertion of DNA to be expressed from said promoter;
wherein said DNA is expressed from said vector in an animal cell.
|
|
Claim 10
A promoter consisting of:
a) a plurality of at least five glucocorticoid response elements (GREs),
b) a TATA box, and
c) an initiator site containing a transcriptional
initiator site located from 20 to 50 bases from said TATA box,
said promoter lacking upstream elements which bind nuclear factor I,
wherein said promoter is responsive to ligand-bound glucocorticoid,
progesterone, androgen or mineralocorticoid receptor when transiently
transfected into cells, when stably integrated within a genome, or when stably
propagated in an episomal vector.
|
2. Promoters based on estrogen receptor (ER)
Patent applications filed by The Rockefeller University
Actual pending independent claims
| EP 1232273 A2 |
|
Claim 1
A method for selecting transgenic plants
comprising a silent selectable marker wherein said method comprises the steps
of:
a) transforming a plant cell with a vector wherein said vector comprises DNA
encoding a regulatory region of an estrogen receptor and
further wherein said vector comprises a gene which promotes shoot formation,
wherein said gene is under the control of an inducible promoter;
b)
growing said plant cells in the absence of a plant hormone but in the presence
of an inducer of said inducible promoter; and
c) excising shoots which
develop, wherein said shoots can grow into transgenic plants when grown in the
absence of said inducer.
|
|
Claim 14
A method for inducing plant somatic embryo
formation comprising the steps of:
a) transforming a plant cell with a
vector encoding a gene which promotes somatic embryogenesis, wherein said gene
is under the control of an inducible promoter; and
b) growing said plant
cells in the absence of a plant hormone but in the presence of an inducer of
said inducible promoter, wherein somatic embryos will develop.
|
|
Claim 26
A method for selecting transgenic plants
wherein said method comprises growing a transgenic plant, comprising an
antibiotic resistance gene under the control of a promoter comprising DNA
encoding a regulatory domain of an estrogen receptor inducible by 17-estradiol
or 4-hydroxyl tamoxifen, in the presence of an antibi otic, wherein said
antibiotic is one to which resistance is conferred by said antibiotic resistance
gene, and in the presence of 17- estradiol or 4-hydroxyl tamoxifen.
|
|
Claim 27
A method for selecting transgenic plants
wherein said method comprises growing a transgenic plant, comprising a herbicide
resistance gene wherein said herbicide resistance gene is under the control of a
promoter comprising DNA encoding a regulatory domain of an estrogen receptor
inducible by 17-estradiol or 4-hydroxyl tamoxifen, in the presence of a
herbicide, wherein said herbicide is one to which resistance is conferred by
said herbicide resistance gene, and in the presence of 17-estradiol or
4-hydroxyl tamoxifen.
|
| EP 1242604 A2 |
|
Claim 1
A method for selecting transgenic lettuce
plants comprising a silent selectable marker wherein said method comprises the
steps of : a) transforming lettuce root cells with a vector wherein said vector
comprises a gene selected from the group consisting of an ipt gene, a CKI1 gene,
a gene from the knotted family, and a gene the expression of which is capable of
promoting shoot regeneration, wherein said gene is under the control of an
inducible promoter ; b) growing said lettuce root cells to allow shoot
development ; and c) excising shoots which develop from plants having a shooty
phenotype, wherein said shoots can grow into normal transgenic plants when grown
in the absence of said inducer.
|
|
Claim 11
A vector comprising a chemically inducible
promoter wherein said vector comprises DNA encoding an estrogen receptor.
|
|
Claim 25
A vector comprising:
i) a constitutive promoter,
ii) DNA encoding a DNA binding domain of
bacterial repressor LexA,
iii) DNA encoding a transactivating domain of
VP16,
iv) DNA encoding an estrogen receptor, and
v) one or more LexA
binding sites.
|
|
Claim 26
A nucleic acid comprising a chemically
inducible promoter wherein said nucleic acid further comprises DNA encoding an
estrogen receptor.
|
|
Claim 41
A nucleic acid comprising i) a constitutive
promoter, ii) DNA encoding a DNA binding domain of bacterial repressor LexA,
iii) DNA encoding a transactivating domain of VP 16, iv) DNA encoding an
estrogen receptor, and v) one or more LexA binding sites.
|
|
Claim 42
A transgenic lettuce plant or transgenic
lettuce plant cell comprising a vector wherein said vector comprises a
chemically inducible promoter.
|
|
Claim 43
A transgenic plant or transgenic plant cell
comprising a vector wherein said vector comprises a chemically inducible
promoter which can be induced by an estrogen.
|
|
Claim 50
A method for making a transgenic plant display
a fluorescent design, a word or words wherein said method comprises the steps of
: a) preparing a transgenic plant which comprises a luciferase gene under the
control of a chemically inducible promoter which is controlled by an estrogen ;
and b) placing a chemical which induces said chemically inducible promoter onto
said transgenic plant in the pattern of the design, word or words which are
desired ; whereby said plant will produce luciferase and will fluoresce in the
pattern in which the chemically inducible promoter was placed onto said
transgenic plant
|
|
Claim 51
A transgenic lettuce plant comprising an
antibiotic resistance gene wherein said antibiotic resistance gene is under the
control of an inducible promoter.
|
|
Claim 52
A transgenic plant comprising an antibiotic
resistance gene wherein said antibiotic resistance gene is under the control of
an inducible promoter, wherein said inducible promoter comprises DNA encoding a
regulatory domain of an estrogen receptor.
|
|
Claim 57
A transgenic plant comprising a herbicide
resistance gene wherein said herb icide resistance gene is under the control of
an inducible promoter, wherein said inducible promoter comprises DNA encoding a
regulatory domain of an estrogen receptor
|
|
Claim 61
An organism or a cell comprising a gene
wherein a natural promoter of said gene is lacking or inoperative and said gene
is under the control of a transgenic inducible promoter
|
|
Claim 74
A method to screen for mutations in a gene of
an organism or cell comprising:
a) preparing an organism or a cell wherein a natural promoter of said gene is
lacking or inoperative and said gene is under the control of a transgenic
inducible promoter; and
b) growing said organism or cell
|
3. Promoters based on ecdysone receptor
Patents granted to Zeneca (now Sygenta)
|
US
6379945
|
|
Claim 1
An isolated or synthetic DNA sequence encoding
a polypeptide selected from the group consisting of:
a) the Heliothis virescens ecdysone steroid receptor shown in SEQ ID
NO: 5;
b) the transactivation domain of the Heliothis virescens
ecdysone steroid receptor shown in amino acids 1-162 of SEQ ID NO: 5;
c)
the DNA binding domain of the Heliothis virescens ecdysone steroid
receptor shown in amino acids 163-228 of SEQ ID NO: 5;
d) the hinge domain
of the Heliothis virescens ecdysone steroid receptor shown in amino
acids 229-326 of SEQ ID NO: 5;
e) the ligand binding domain of the
Heliothis virescens ecdysone steroid receptor shown in amino acids
327-545 of SEQ ID NO: 5;
f) the carboxy terminus of the Heliothis
virescens ecdysone steroid receptor shown in amino acids 546-577 of SEQ ID
NO: 5; and
g) the hinge and ligand binding domains of the Spodoptera
exigua ecdysone steroid receptor shown in SEQ ID NO: 7.
|
|
AU
711391 B2
|
|
Claim 1
DNA comprising the sequence shown in Seq. ID
No. 2, or a DNA sequence which hybridizes to said DNA sequence under high
stringency conditions.
|
|
Claim 2
DNA comprising the sequence shown in Seq. ID
No.3, or a DNA sequence which hybridizes to said DNA sequence under high
stringency conditions.
|
|
Claim 3
DNA comprising the sequence shown in Seq. ID
No. 4, or a DNA sequence which hybridizes to said DNA sequence under high
stringency conditions
|
|
Claim 11
A polypeptide comprising the
Heliothis ecdysone receptor or a func tionally active fragment thereof,
wherein the polypeptide is substantially free from other proteins with which is
ordinarily associated, and which is coded for by the DNA of any one of the
preceding claims
|
|
Claim 12
A polypeptide comprising the amino acid
sequence shown in Seq. ID. No. 4 or any allelic variant or derivative thereof.
|
|
Claim 13
A polypeptide comprising part of the amino
acid sequence shown in Seq. ID. No. 4 or any allelic variant or derivative
thereof, which sequence provides the Heliothis ecdysone
receptor ligand binding domain.
|
|
Claim 14
A polypeptide comprising part of the amino
acid sequence shown in Seq. ID. No. 4 or any allelic variant or derivative
thereof, which sequence provides the Heliothis ecdysone
receptor DNA binding domain.
|
|
Claim 15
A polypeptide comprising part of the amino
acid sequence shown in Seq. ID. No. 4 or any allelic variant or derivative
thereof, which sequence provides the Heliothis ecdysone
receptor transactivation domain.
|
|
Claim 16
A polypeptide comprising part of the amino
acid sequence shown in Seq. ID. No. 4 or any allelic variant or derivative
thereof, which sequence provides the Heliothis ecdysone
receptor hinge domain.
|
|
Claim 17
A polypeptide comprising part of the amino
acid sequence shown in Seq. ID. No. 4 or any allelic variant or derivative
thereof, which sequence provides the Heliothis ecdysone
receptor carboxy terminal domain.
|
|
Claim 19
DNA comprising the sequence shown in Seq. ID.
No. 6, or a DNA sequence which hybridizes to said DNA sequence under high
stringency conditions.
|
Patent applications filed by Syngenta
| EP 828829 A1 |
|
Claim 1
DNA comprising the sequence shown in Seq ID No.
2.
|
|
Claim 2
DNA comprising the sequence shown in Seq ID No.
3.
|
|
Claim 3
DNA comprising the sequence shown in Seq ID No.
4.
|
|
Claim 4
DNA comprising a sequence which shows 60% or
more homology with the sequence shown in Seq ID No 1, 2 or 3.
|
|
Claim 6
DNA which hybridizes to the sequence shown in
Seq. ID No. 2, 3 or 4, and which codes for at least part of the
Heliothis ecdysone receptor.
|
|
Claim 8
DNA comprising part of the sequence shown in
Seq ID No. 2, and which codes for at least part of the Heliothis
ecdysone receptor ligand binding domain.
|
|
Claim 9
DNA comprising part of the sequence shown in
Seq ID No. 3, and which codes for at least part of the Heliothis
ecdysone receptor ligand binding domain.
|
|
Claim 10
DNA comprising part of the sequence shown in
Seq ID No. 4, and which codes for at least part of the Heliothis
ecdysone receptor ligand binding domain.
|
|
Claim 15
DNA comprising part of the sequence shown in
Seq ID No. 2, and which codes for at least part of the Heliothis
ecdysone receptor DNA binding domain.
|
|
Claim 16
DNA comprising part of the sequence shown in
Seq ID No. 3, and which codes for at least part of the Heliothis
ecdysone receptor DNA binding domain.
|
|
Claim 17
DNA comprising part of the sequence shown in
Seq ID No. 4, and which codes for at least part of the Heliothis
ecdysone receptor DNA binding domain.
|
|
Claim 22
DNA comprising part of the sequence shown in
Seq ID No. 2, and which codes for at least part of the Heliothis
ecdysone receptor transactivation domain.
|
|
Claim 23
DNA comprising part of the sequence shown in
Seq ID No. 3, and which codes for at least part of the Heliothis
ecdysone receptor transactivation domain.
|
|
Claim 24
DNA comprising part of the sequence shown in
Seq ID No. 4, and which codes for at least part of the Heliothis
ecdysone receptor transactivation domain.
|
|
Claim 29
DNA comprising part of the sequence shown in
Seq ID No. 2, and which codes for at least part of the Heliothis
ecdysone receptor hinge domain.
|
|
Claim 30
DNA comprising part of the sequence shown in
Seq ID No. 3, and which codes for at least part of the Heliothis
ecdysone receptor hinge domain.
|
|
Claim 31
DNA comprising part of the sequence shown in
Seq ID No. 4, and which codes for at least part of the Heliothis
ecdysone receptor hinge domain.
|
|
Claim 36
DNA having part of the sequence shown in Seq
ID No. 2, and which codes for at least part of the Heliothis ecdysone
receptor carboxy terminal region.
|
|
Claim 37
DNA having part of the sequence shown in Seq
ID No. 3, and. which codes for at least part of the Heliothis ecdysone
receptor carboxy terminal region.
|
|
Claim 38
DNA having part of the sequence shown in Seq
ID No. 4, and which codes for at least part of the Heliothis ecdysone
receptor carboxy terminal region.
|
|
Claim 44
A polypeptide comprising the amino acid
sequence shown in Seq ID No. 4 or any allelic variant or derivative thereof.
|
|
Claim 45
A polypeptide comprising part of the amino
acid sequence shown in Seq ID No. 4 or any allelic variant or derivative
thereof, which sequence provides the Heliothis ecdysone
receptor ligand binding domain.
|
|
Claim 46
A polypeptide comprising part of the amino
acid sequence shown in Seq ID No. 4 or any allelic variant or derivative
thereof, which sequence provides the Heliothis ecdysone
receptor DNA binding domain.
|
|
Claim 47
A polypeptide comprising part of the amino
acid sequence shown in Seq ID No. 4 or any allelic variant or derivative
thereof, which sequence provides the Heliothis ecdysone
receptor transactivation domain.
|
|
Claim 48
A polypeptide comprising part of the amino
acid sequence shown in Seq ID No. 4 or any allelic variant or derivative
thereof, which sequence provides the Heliothis ecdysone
receptor hinge domain.
|
|
Claim 49
A polypeptide comprising part of the amino
acid sequence shown in Seq ID No. 4 or any allelic variant or derivative
thereof, which sequence provides the Heliothis ecdysone
receptor carboxy terminal region.
|
|
Claim 51
DNA comprising the sequence shown in Seq ID
No. 6.
|
|
Claim 52
DNA comprising a sequence which shows 60% or
more homology with the sequence shown in Seq ID No. 6.
|
|
Claim 54
DNA which hybridizes to the DNA sequence shown
in Seq ID No. 6 and which codes for at least part of Spodoptera
ecdysone receptor.
|
|
Claim 56
DNA comprising part of the sequence shown in
Seq ID No. 6, and which codes for at least part of the Spodoptera
ecdysone receptor ligand binding domain.
|
|
Claim 61
DNA comprising part of the sequence shown in
Seq ID No. 6, and which codes for at least part of the Spodoptera
ecdysone receptor hinge domain.
|
| CA 2219121 AA |
|
Claim 1
DNA comprising the sequence shown in Seq. ID
No. 2, or a sequence which shows homology thereto, or a sequence which
hybridizes to the sequence shown in Seq. ID No. 2.
|
|
Claim 2
DNA comprising the sequence shown in Seq. ID
No.3, or a sequence which shows homology thereto, or a sequence which hybridizes
to the sequence shown in Seq. ID No. 3.
|
|
Claim 3
DNA comprising the sequence shown in Seq. ID
No. 4, or a sequence which shows homology thereto, or a sequence which
hybridizes to the sequence shown in Seq. ID No. 4.
|
|
Claim 14
A polypeptide comprising the amino acid
sequence shown in Seq. ID. No. 4 or any allelic variant or derivative thereof.
|
|
Claim 15
A polypeptide comprising part of the amino
acid sequence shown in Seq. ID. No. 4 or any allelic variant or derivative
thereof, which sequence provides the Heliothis ecdysone
receptor ligand binding domain.
|
|
Claim16
A polypeptide comprising part of the amino acid sequence shown
in Seq. ID. No. 4 or any allelic variant or derivative thereof, which sequence
provides the Heliothis ecdysone receptor DNA binding
domain.
|
|
Claim 17
A polypeptide comprising part of the amino
acid sequence shown in Seq. ID. No. 4 or any allelic variant or derivative
thereof, which sequence provides the Heliothis ecdysone
receptor transactivation domain.
|
|
Claim 18
A polypeptide comprising part of the amino
acid sequence shown in Seq. ID. No. 4 or any allelic variant or derivative
thereof, which sequence provides the Heliothis ecdysone
receptor hinge domain.
|
|
Claim 19
A polypeptide comprising part of the amino
acid sequence shown in Seq. ID. No. 4 or any allelic variant or derivative
thereof, which sequence provides the Heliothis ecdysone
receptor carboxy terminal domain.
|
|
Claim 21
DNA comprising the sequence shown in Seq. ID.
No. 6, or a sequence which shows homology thereto, or a sequence which
hybridizes to the sequence shown in Seq. ID. No. 6.
|
|
Claim 23
DNA comprising a sequence which shows homology
in the range of 65% to 99% to the sequence shown in Seq. ID. No. 6.
|
Patent application filed by Pioneer Hi-Bred
| EP 1112360 A1 |
|
Claim 1
An isolated nucleic acid sequence that encodes
an insect protein from a Pyralidae species, wherein said nucleotide sequence is
selected from the group consisting of:
a) a nucleotide sequence comprising a sequence encoding an ecdysone receptor
or Ultraspiracle;
b) a nucleotide sequence comprising the sequence set
forth in SEQ ID NO: 1 or 3;
c) a nucleotide sequence encoding a
polypeptide comprising the amino acid sequence set forth in SEQ ID NO: 2 or 4;
d) a nucleotide sequence that hybridizes under stringent conditions to a
sequence of a), b), or c).
|
|
Claim 2
An isolated polypeptide from a Pyrilidae
species, wherein said polypeptide is selected from the group consisting of.
a) a polypeptide sequence comprising an Ecdysone receptor or Ultraspiracle;
b) a polypeptide comprising an amino acid sequence set forth in 20 SEQ ID
NO: 2 or 4;
c) a polypeptide encoded by a nucleotide sequence comprising
the sequence set forth in SEQ ID NO: 1 or 3;
d) a polypeptide encoded by a
nucleotide sequence that hybridizes under stringent conditions to a nucleotide
sequence comprising the sequence set forth in SEQ ID NO: 1 or 3.
|
|
Claim 3
A method of selectively inducing gene
expression of a protein of interest in a plant, said method comprising:
a) stably incorporating into the genome of said plant an expression cassette,
said expression cassette comprising a promoter operably linked to a nucleotide
sequence encoding an Ecdysone receptor, wherein said nucleotide sequence
encoding the Ecdysone receptor is selected from the group consisting of.:
i) a nucleotide sequence from a Pyrilidae species comprising a sequence
encoding an Ecdysone receptor;
ii) a nucleotide sequence comprising the
sequence set forth in SEQ ID NO: 1;
iii) a nucleotide sequence encoding a
polypeptide comprising the amino acid sequence set forth in SEQ ID NO: 2;
iv) a nucleotide sequence that hybridizes under stringent conditions to a
sequence of i), ii), or iii);
b) further stably incorporating into the genome of said plant a second
expression cassette,
wherein said expression cassette comprises a
transcriptional regulatory region operably linked to a nucleotide sequence
encoding said protein of interest, and
wherein said transcriptional
regulatory region is activated by the ligand-receptor complex;
c)
contacting said plant with a ligand which complexes with said receptor,
wherein said receptor-ligand complex interacts with the transcription regulatory
region and induces gene expression of the protein of interest.
|
|
Claim 23
An expression vector comprising a promoter
operably linked to a nucleotide sequence encoding an Ecdysone receptor wherein
said nucleotide sequence encoding the Ecdysone receptor is selected from the
group consisting of:
a) a nucleotide sequence from a Pyrilidae species comprising a sequence
encoding an Ecdysone receptor;
b) a nucleotide sequence comprising the
sequence set forth in SEQ ID NO: 1;
c) a nucleotide sequence encoding a
polypeptide comprising the amino acid sequence set forth in SEQ ID NO: 2;
d) a nucleotide sequence that hybridizes under stringent conditions to a
sequence of a), b), or c).
|
|
Claim 25
An expression vector comprising a promoter
operably linked to a nucleotide sequence encoding ultraspiracle, and said
nucleotide sequence encoding ultraspiracle is selected from the group consisting
of:
a) a nucleotide sequence comprising the sequence set forth in SEQ ID NO: 3;
b) a nucleotide sequence encoding a polypeptide comprising the amino acid
sequence set forth in SEQ ID NO: 4;
c) a nucleotide sequence that
hybridizes under stringent conditions to a sequence of a) or b).
|
Patents on yeast copper-responsive promoter
Patent granted to Genentech
|
US
4940661
|
|
Claim 1
A vector comprising a selection gene, yeast
origin of replication or autonomously replicating sequence and DNA encoding a
eukaryotic polypeptide other than yeast chelatin, said DNA encoding a eukaryotic
polypeptide that is under the control of:
a) a yeast chelatin promoter,
b) a yeast chelatin transcription control
sequence, or
c) a metal ion regulatory region of the yeast chelatin
transcription control sequence, which is free of the yeast chelatin promoter,
whereby the vector is replicable in a suitable host.
|
|
Claim 14
An isolated DNA sequence comprising a yeast
chelatin metal ion regulatory region free of the yeast chelatin promoter and
free of DNA encoding yeast che latin.
|
Patent on a mouse metallothionein promoter
Patent granted to University Patents Inc.
|
US
4579821
|
|
Claim 1
A process for subjecting the transcription of a
selected DNA sequence to external control under given environmental conditions
which comprises the steps of:
A) providing a selected isolated structural gene that is transcriptionally
responsive to a mouse metallothionein-I promoter/regulator DNA sequence under
the given environmental conditions; and
B) operatively fusing the selected
structural gene with said promoter/regulator DNA sequence.
|
|
Claim 2
In the genetic engineering process for securing
transcription and expression of a selected isolated structural gene sequence in
a mammalian host cell wherein said selected structural gene is stably
incorporated as a chromosomal or extrachromosomal constituent of the host, the
improvement comprising the step of:
operatively fusing with said selected structural gene sequence a mouse
metallothionein-I promoter/regulator DNA sequence, which is responsive to
environmental variations within the host cell in the concentration of ions of
metal.
|
|
Claim 3
A fusion gene product, suitable for use in
genetic transformation of a mammalian host cell, said product comprising:
a non mouse metallothionein structural gene sequence to be incorporated in
said host cell operatively fused with a mouse metallothionein-I
promoter/regulator DNA sequence.
|
Patent on a human metallothionein promoter
Patent granted to the University of California (Berkeley)
|
US
4601978
|
|
Claim 1
A DNA sequence of less than 500 base pairs,
said DNA sequence comprising the human MT-II transcriptional regulatory system
further comprising the transcription initiation sequence.
|
|
Claim 5
A DNA construct capable of regulated expression
of an inserted gene in a mammalian host, said construct comprising an
extrachromosomal replication system recognized by a mammalian host, which
replication system has been joined to a human MT-II gene inducible regulatory
system in vitro, said construct having DNA sequence(s) defining at least one
restriction site within the transcriptional control of the regulatory region for
insertion of the gene.
|
|
Claim 10
A DNA construct useful for expression of a
structural gene to produce a polypeptide in a mammalian host, said vector
comprising (a) a replicon from bovine papilloma virus and (b) a regulatory
system from a human MT-II gene comprising a promoter and a terminator and having
at least one restriction site downstream from the promoter in the direction of
transcription.
|
Patents granted to Novartis (now Syngenta)
|
US
5654414
|
|
Claim 1
A nucleic acid promoter fragment isolated from
the 5' flanking region upstream of the coding region of a cucumber
chitinase/lysozyme gene that is inducible by application of
benzo-1,2,3-thiadiazoles.
|
|
US
5689044
|
|
Claim 1
A chemically inducible nucleic acid promoter
fragment isolated from the 5' flanking region upstream of the coding region of a
tobacco PR-1a gene, wherein said promoter fragment comprises a nucleotide
fragment of at least 603-bp adjacent to the coding region of said tobacco PR-1a
gene, wherein said promoter fragment is inducible by application of a
benzo-1,2,3-thiadiazole, an isonicotinic acid compound, or a salicylic acid
compound.
|
|
Claim 3
A chemically inducible nucleic acid promoter
fragment isolated from the 5' flanking region upstream of the coding region of
an Arabidopsis PR-1 gene, wherein the coding region of said Arabidopsis PR-1
gene comprises the DNA sequence set forth in SEQ ID NO:33 or a DNA sequence
which would encode the protein encoded by SEQ ID NO:33,
wherein said
promoter fragment is inducible by application of a ben zo-1,2,3-thiadiazole, an
isonicotinic acid compound, or a salicylic acid compound.
|
|
US
5789214
|
|
Claim 1
A method of inducing gene transcription in a
plant or plant tissue, comprising the steps of:
A) transforming said plant or plant tissue, each with a chimeric gene
comprising:
(i) a chemically inducible nucleic acid promoter fragment of
at least 603-bp isolated from the 5' flanking region adjacent the coding region
of a tobacco PR-1a gene, and
(ii) a coding sequence of interest
operatively linked to said promoter fragment; and
B) exposing said
transgenic plant or plant tissue to a benzo-1,2,3-thiadiazole, an isonicotinic
acid compound, or a salicylic acid compound, whereby transcription of said
coding sequence of interest is induced in said plant or plant tissue.
|
|
Claim 8
A method of inducing gene transcription in a
plant or plant tissue, comprising the steps of:
A) transforming said plant or plant tissue, each with a chimeric gene
comprising:
(i) a chemically inducible nucleic acid promoter fragment
isolated from the 5' flanking region adjacent the coding region of an
Arabidopsis PR-1 gene, wherein said Arabidopsis PR-1 gene comprises a DNA
sequence that specifically hybridizes to SEQ ID NO:33 or wherein said
Arabidopsis PR-1 gene comprises a DNA sequence that encodes the protein encoded
by SEQ ID NO:33, and
(ii) a coding sequence of interest operatively linked
to said promoter fragment; and
B) exposing said transgenic plant or plant
tissue to a benzo-1,2,3-thiadiazole, an isonicotinic acid compound, or a
salicylic acid compound, whereby transcription of said coding sequence of
interest is induced in said plant or plant tissue.
|
|
AU
708850 B2
|
|
Claim 1
An isolated DNA molecule comprising a
nucleotide sequence selected from the following group:
a) a full-length chemically inducible promoter fragment comprising
nucleotides 1 through 4258 of SEQ ID NO: 1;
b) an 815-bp long chemically
inducible promoter fragment comprising nucleotides 3444 through 4258 of SEQ ID
NO: 1; and
c) a 698-bp long chemically inducible promoter fragment
comprising nucleotides 3561 through 4258 of SEQ ID NO: 1.
|
|
Claim 14
An isolated DNA molecule involved in
inducibility of a chemically inducible promoter selected from the following
group:
a) LS4 comprising nucleotides 3584 through 3593 of SEQ ID NO: 1;
b) LS7
comprising nucleotides 3614 through 3623 of SEQ ID NO: 1;
c) LS 10
comprising nucleotides 3644 through 3653 of SEQ ID NO: 1; and
d) a region
spanning LS7-LS 1 0 and comprising nucleotides 3614 through 3653 of SEQ ID NO:
1.
|
Patent granted to Pioneer Hi-Bred
|
US
6429362
|
|
Claim 1
An isolated promoter comprising a nucleotide
sequence that initiates transcription in a plant cell, wherein said nucleotide
sequence is selected from the group consisting of:
a) a nucleotide sequence comprising the sequence set forth in SEQ ID NO: 3 or
4; and
b) a nucleotide sequence comprising the plant promoter sequence
deposited in the plasmid designated as ATCC Accession No. 207139 or 207131.
|
|
Claim 5
A method for driving expression of a
heterologous nucleotide sequence in a plant, said method comprising the steps
of:
a) transforming a plant cell with an expression cassette comprising the
heterologous nucleotide sequence operably linked to a promoter that initiates
transcription in a plant cell, wherein said promoter is selected from the group
consisting of:
i) a nucleotide sequence comprising the sequence set forth
in SEQ ID NO: 3 or 4; and
ii) a nucleotide sequence comprising the plant
promoter sequence deposited in the plasmid designated as ATCC Accession No.
207139 or 207131; and
b) regenerating a stably transformed plant from said
plant cell.
|
|
Claim 9
A plant cell transformed with a DNA construct
comprising a heterologous nucleotide sequence operably linked to a promoter that
initiates transcription in said plant cell, wherein said promoter comprises a
nucleotide sequence selected from the group consisting of:
a) a nucleotide sequence comprising the sequence set forth in SEQ ID NO: 3 or
4; and
b) a nucleotide sequence comprising the plant promoter sequence
deposited in the plasmid designated as ATCC Accession No. 207139 or 207131.
|
|
Claim 13
A plant stably transformed with a DNA
construct comprising a heterologous nucleotide sequence operably linked to a
promoter that initiates transcription in a plant cell, wherein said promoter
comprises a nucleotide sequence selected from the group consisting of:
a) a nucleotide sequence comprising the sequence set forth in SEQ ID NO: 3 or
4; and
b) a nucleotide sequence comprising the plant promoter sequence
deposited in the plasmid designated as ATCC Accession No. 207139 or 207131.
|
Patent application filed by Pioneer Hi-Bred
| EP 1056862 A1 |
|
Claim 1
An isolated nucleic acid molecule having a
nucleotide sequence for a promoter that is capable of initiating transcription
in a plant cell,
wherein said nucleotide sequence is selected from the
group consisting of:
a) a nucleotide sequence comprising the sequence set forth in SEQ ID NO: 1,
2, 3, 4, or 5;
b) a nucleotide sequence selected from the group consisting
of sequences deposited as ATCC Accession No. _
c) a nucleotide sequence
comprising at least 40 contiguous 10 nucleotides of the sequence set forth in
SEQ ID NO: 1, 2, 3, 4, or 5; and
d) a nucleotide sequence that hybridizes
under stringent conditions to a sequence of a), b), or c).
|
|
Claim 5
A method for inducing expression of a
heterologous nucleotide sequence in a plant, said method comprising:
A) transforming a plant cell with a DNA construct comprising said
heterologous nucleotide sequence operably linked to a promoter that is capable
of initiating transcription in a plant cell in response to a stimulus,
B)
regenerating a stably transformed plant from said plant cell, and
C)
exposing said plant to said stimulus, wherein said promoter comprises a
nucleotide sequence selected from the group consisting of:
a) a nucleotide sequence comprising the sequence set forth in 30 SEQ ID NO:
1, 2, 3, or 4;
b) a nucleotide sequence selected from the group consisting
of sequences deposited as ATCC Accession No._
c) a nucleotide sequence
comprising at least 40 contiguous 5 nucleotides of the sequence set forth in SEQ
ID NO: 1, 2, 3, or 4; and
d) a nucleotide sequence that hybridizes under
stringent conditions to a sequence of a), b), or c).
|
|
Claim 9
A method for constitutively expressing a
heterologous nucleotide sequence in a plant, said method comprising:
A) transforming a plant cell with a DNA construct comprising said
heterologous nucleotide sequence operably linked to a promoter that is capable
of initiating constitutive transcription in a plant cell and
B)
regenerating a stably transformed plant from said plant cell,
wherein said
promoter comprises a nucleotide sequence selected from the group consisting of:
a) a nucleotide sequence comprising the sequence set forth in SEQ ID NO: 5;
b) a nucleotide sequence selected from the group consisting of sequences
deposited as ATCC Accession No._
c) a nucleotide sequence comprising at
least 40 contiguous nucleotides of the sequence set forth in SEQ ID NO: 5; and
d) a nucleotide sequence that hybridizes under stringent conditions to a
sequence of a), b), or c).
|
|
Claim 13
A plant cell stably transformed with a DNA
construct comprising a heterologous nucleotide sequence operably linked to a
promoter that is capable of initiating transcription in said plant cell,
wherein said promoter comprises a nucleotide sequence selected from the group
consisting of :
a) a nucleotide sequence comprising the sequence set forth in SEQ ID NO: 1,
2, 3, 4, or 5;
b) a nucleotide sequence selected from the group consisting
of sequences deposited as ATCC Accession No._
c) a nucleotide sequence
comprising at least 40 contiguous nucleotides of the sequence set forth in SEQ
ID NO: 1, 2, 3, 4, or 5; and
d) a nucleotide sequence that hybridizes under
stringent conditions to a sequence of a), b), or c).
|
|
Claim 17
A plant stably transformed with a DNA
construct comprising a heterologous nucleotide sequence operably linked to a
promoter that is capable of initiating transcription in a plant cell,
wherein said promoter comprises a nucleotide sequence selected from the group
consisting of:
a) a nucleotide sequence comprising the sequence set forth in SEQ ID NO: 1,
2, 3, 4, or 5;
b) a nucleotide sequence selected from the group consisting
of sequences deposited as ATCC Accession No. _
c) a nucleotide sequence
comprising at least 40 contiguous nucleotides of the sequence set forth in SEQ
ID NO: 1, 2, 3, 4, or 5; and
d) a nucleotide sequence that hybridizes
under stringent conditions to a sequence of a), b), or c).
|
|
Claim 22
An isolated nucleic acid molecule having a
nucleotide sequence selected from the group consisting of:
a) a nucleotide sequence comprising the sequence set forth in SEQ ID NO: 6,
8, 10, or 14;
b) a nucleotide sequence selected from the group consisting
of sequences deposited as ATCC Accession No._
c) a nucleotide sequence
encoding a polypeptide comprising the amino acid sequence set forth in SEQ ID
NO: 7, 9, 11, or 15;
d) a nucleotide sequence comprising at least 16
contiguous nucleotides of a sequence of a), b), or c); and
e) a nucleotide
sequence that hybridizes under stringent conditions to a sequence of a), b), c),
or d).
|
|
Claim 26
A method for creating or enhancing disease
resistance in a plant, said method comprising:
A) transforming said plant with a DNA construct comprising a PR-1 sequence
operably linked to a promoter that drives expression of a coding sequence in a
plant cell and
B) regenerating stably transformed plants,
wherein
said PR-1 sequence is selected from the group consisting of:
a) a nucleotide sequence comprising the sequence set forth in SEQ ID NO: 6,
8, 10, or 14;
b) a nucleotide sequence selected from the group consisting
of sequences deposited as ATCC Accession No._
c) a nucleotide sequence
encoding a polypeptide comprising the amino acid sequence set forth in SEQ ID
NO: 7, 9, 11, or 15;
d) a nucleotide sequence comprising at least 16
contiguous nucleotides of a sequence of a), b), or c); and
e) a nucleotide
sequence that hybridizes under stringent conditions to a se quence of a), b),
c), or d).
|
|
Claim 33
A plant cell stably transformed with a DNA
construct comprising a PR-1 sequence operably linked to a promoter that drives
expression of a coding sequence in a plant cell, wherein said PR-1 sequence is
selected from the group consisting of:
a) a nucleotide sequence comprising the sequence set forth in SEQ ID NO: 6,
8, 10, or 14;
b) a nucleotide sequence selected from the group consisting
of sequences deposited as ATCC Accession No._
c) a nucleotide sequence
encoding a polypeptide comprising the amino acid sequence set forth in SEQ ID
NO: 7, 9, 11, or 15;
d) a nucleotide sequence comprising at least 16
contiguous nucleotides of a sequence of a), b), or c); and
e) a nucleotide
sequence that hybridizes under stringent conditions to a sequence of a), b), c),
or d).
|
|
Claim 34
A plant stably transformed with a DNA
construct comprising a PR-1 sequence operably linked to a promoter that drives
expression of a coding sequence in a plant cell, wherein said PR-1 sequence is
selected from the group consisting of:
a) a nucleotide sequence comprising the sequence set forth in SEQ ID NO: 6,
8, 10, or 14;
b) a nucleotide sequence selected from the group consisting
of sequences deposited as ATCC Accession No._
c) a nucleotide sequence
encoding a polypeptide comprising the amino acid sequence set forth in SEQ ID
NO: 7, 9, 11, or 15;
d) a nucleotide sequence comprising at least 16
contiguous nucleotides of a sequence of a), b), or c); and
e) a nucleotide
sequence that hybridizes under stringent conditions to a sequence of a), b), c),
or d).
|
Heat-regulated
Actual granted independent claims
Patents granted to Mycogen Plant Science
|
US
5447858
|
|
Claim 1
A recombinant heat shock gene comprising a
plant heat shock promoter and a heterologous structural gene whose expression is
controllable thereby,
wherein said promoter comprises the consensus
nucleotide sequence 5'-C-T-X-G-A-A-X-X-T-A-C-X-X-X-3',
where X is A,T,C or
G.
|
|
Claim 2
A recombinant heat shock gene comprising a
plant heat shock promoter and a heterologous structural gene whose expression is
controlled thereby,
wherein said promoter comprises the consensus
nucleotide sequence 5'-C-T-S-G-A-A-M-R-T-A-C-W-M-K-3',
where S is C or G;
M is A or C; R is A or G; W is A or T; and K is T or G.
|
|
Claim 8
A vector comprising a fragment of DNA capable
of functioning as a plant heat shock promoter wherein said fragment of DNA
comprises the consensus nucleotide sequence 5'-C-T-X-G-A-A-X-X-T-A-C-X-X-X-3',
where X is A, T, C or G.
|
|
Claim 9
A vector comprising a fragment of DNA capable
of functioning as a plant heat shock promoter wherein said fragment comprises
the consensus nucleotide sequence of 5'-C-T-S-G-A-A-M-R-T-A-C-W-M-K-3',
where S is C or G; M is A or C; R is A or G; W is A or T; and K is T or G.
|
|
Claim 19
A bacterial strain containing therein
recombinant DNA comprising:
a) a fragment of DNA capable of controlling heat shock expression of a gene
in a plant; and
b) a structural gene oriented with respect to said fragment
of DNA so as to be expressible under the control thereof
wherein said
fragment of DNA capable of controlling heat shock expression of a gene in a
plant comprises the consensus nucleotide sequence
5'-C-T-X-G-A-A-X-X-T-A-C-X-X-X-3',
where X is A, T, C or G.
|
|
Claim 20
A bacterial strain containing therein
recombinant DNA comprising:
a) a fragment of DNA capable of controlling heat shock expression of a gene
in a plant; and
b) a structural gene oriented with respect to said fragment
of DNA so as to be expressible under the control thereof
wherein the
fragment of DNA capable of controlling heat shock expression of a gene in a
plant comprises the consensus nucleotide sequence
5'-C-T-S-G-A-A-M-R-T-A-C-W-M-K-3',
where S is C or G; M is A or C; R is A
or G; W is A or T; and K is T or G.
|
|
Claim 34
A method for recognizing a plant cell
containing recombinant DNA comprising the steps of:
1) transferring into said plant cell recombinant DNA comprising:
a) a
heat shock promoter which comprises the consensus sequence
5'-C-T-X-G-A-A-X-X-T-A-C-X-X-X-3', where X is A, T, C, or G;
b) a
transformation recognition gene under the control of said heat shock promoter
capable of causing an observable or detectable reaction when expressed;
2) applying a stress to said plant cell in which it is desired to recognize
those containing recombinant DNA, which stress is capable of inducing a response
in said heat shock promoter; and
3) observing or detecting the reaction
caused by expression of said transformation recognition gene under the control
of said heat shock promoter to recognize said plant cell containing recombinant
DNA including said tra nsformation recognition gene.
|
|
Claim 36
A method for recognizing a plant cell
containing recombinant DNA comprising the steps of:
1) transferring into said plant cell recombinant DNA comprising:
a) a
heat shock promoter which comprises the consensus sequence
5'-C-T-S-G-A-A-M-R-T-A-C-W-M-K-3',
where S is C or G; M is A or C; R is A
or G; W is A or T; and K is T or G;
b) a transformation recognition gene
under the control of said heat shock promoter capable of causing an observable
or detectable reaction when expressed;
2) applying a stress to said plant
cell in which it is desired to recognize those containing recombinant DNA, which
stress is capable of inducing a response in said heat shock promoter; and
3) observing or detecting the reaction caused by expression of said
transformation recognition gene under the control of said heat shock promoter to
recognize said plant cell containing recombinant DNA including said
transformation recognition gene.
|
|
EP
159884 B1
|
|
Claim 1
A recombinant DNA plasmid comprising:
a) a vector,
b) a T-DNA fragment of a Ti-plasmid from an
Agrobacterium strain, which T-DNA fragment is functional for insertion
into a plant genome, and
c) a fragment of plant DNA which is capable of
controlling gene expression in response to heat shock, and which fragment is
inserted into said T-DNA fragment such that said T-DNA fragment retains
functionality for insertion into a plant genome,
wherein said fragment of
plant DNA comprises a plant heat shock promoter comprising the consensus
nucleotide sequence 5'-C-T-X-G-A-A-X-X-T- A-C-X-X-X-3', wherein X is A, T, C or
G.
|
|
Claim 17
A method for modifying a plant genome so as to
allow expression of a structural gene under control of a
soybean heat shock gene promoter fragment, comprising the steps
of:
a) isolating a soybean heat shock gene promoter fragment
comprising the consensus nucleotide sequence 5'-C-T-X-G-A-A-X-X-T-A-C-X-X-X-3',
wherein X is A, T, C or G,
b) cloning said soybean heat shock gene
promoter fragment into a T-DNA shuttle vector producing a recombinant DNA
plasmid,
c) isolating a DNA fragment carrying foreign structural genes or
soybean genes not naturally expressed under the regulatory control of a soybean
heat shock gene promoter, and inserting said DNA fragment into said recombinant
DNA plasmid at a position on the X-side of said soybean heat shock gene promoter
producing a heat shock expression plasmid,
wherein said DNA fragment is
oriented with respect to said soybean heat shock gene promoter as to be
expressible under control thereof,
d) transforming said heat shock
expression plasmid into a first bacterial strain capable of supporting
replication of said heat shock expression plasmid,
e) mixing said
bacterial strain capable of supporting replication of said heat shock expression
plasmid with a second bacterial strain carrying a helper plasmid capable of
transporting said heat shock expression plasmid into an Agrobacterium
strain incapable of supporting replication of said heat shock expression
plasmid, said Agrobacterium strain carrying a resident plasmid,
f) selecting for recombination between said heat shock expression plasmid and
said resident plasmid giving a recombinant resident plasmid,
g) infecting a
plant or a plant cell culture with said Agrobacterium strain containing
and replicating said recombinant resident plasmid, and
h) selecting a
plant or a plant cell culture comprising plant cells containing said foreign
structural genes or said soybean genes under control of said soybean heat shock
gene promoter transferred from said recombinant resident plasmid to said plant
cells,
said foreign structural genes or said soybean genes being expressed
following heat shock treatment or other stress treatment.
|
| CA 1338010 |
|
Claims 1, 8, and 19 are worded exactly the same as claims 1, 8 and 19
of the United States patent.
|
|
Claim 34
A method for obtaining a transient increase in
expression level of a heterologous structural gene in a plant comprising:
1) inserting into said plant genome a DNA fragment comprising a promoter
capable of inducing a heat shock response in a plant exposed to stress,
which promoter comprises the consensus nucleotide sequence
5'-C-T-X-G-A-A-X-X-T-A-C-X-X-X-5', where X is A, T, C or G,
said promoter
being combined with said structural gene such that said structural gene is
expressed under the control of said promoter, followed by 2) exposing said plant
to stress such that said promoter responds thereby obtaining a transient
increase in expression level of said structural gene in said plant.
|
|
Claim 50
A method for controlling the expression of a
structural gene in a plant comprising the steps of:
1) transforming said plant with a DNA fragment comprising a DNA promoter
sequence capable of controlling heat shock expression in a plant and a
structural gene, wherein said DNA promoter sequence comprises the consensus
nucleotide sequence 5'-C-T-X-G-A-A-X-X-T-A-C-X-X-X-3',
where X is A, T, C
or G and said structural gene is oriented with respect to said DNA promoter
sequence so as to be controllable thereby;
2) applying stress to said
plant so as to activate said DNA promoter sequence and thereby cause the
expression of said structural gene.
|
|
Claim 54
It is worded exactly the same as claim 34 of the
United States patent.
|
Patent granted to the U.S. Department of Health and Human Services
|
AU
732872 B2 * |
|
Claim 1
A method for the spatial and temporal control
of the expression of a gene of interest within a preselected discrete region of
a cell mass or multicellular organism, comprising selectively heating a
preselected discrete region of a cell mass or multicellular organism that
includes cells that contain a genetically engineered gene of interest operably
linked to a heat-inducible promoter, thereby inducing the expression of said
gene of interest in the cells that are selectively heated.
|
|
Claim 9
A method of providing a therapeutic
protein to selected cells in an animal, comprising the
steps of:
A) introducing into cells of an animal a DNA molecule having a heat shock
promoter sequence operably linked to and exerting regulatory control over a
sequence encoding a therapeutic protein, and
B) activating said heat shock
promoter sequence through the application of a focused ultrasound so that said
DNA segment expresses a therapeutically effective amount of said therapeutic
protein.
|
|
* Independent claims as filed of the European patent
application EP 922110 A2 are the same as the independent,
granted claims of the Australian patent.
|
Patent granted to The General Hospital Corporation
| CA 1324097 |
|
Claim 1
A method for high level production of a
polypeptide or protein in a transformed host cell comprising:
a) transforming a host cell with a structural gene encoding for a polypeptide
or protein, under the control of an inducible heat shock promoter;
b)
amplifying the copy number of said structural gene in said transformed host
cells using an amplification system under the control of a promoter other than
an inducible heat shock promoter;
c) inducing said inducible promoter by
heat shock to said transformed host cells at a temperature and for a time
sufficient to transcribe said structural gene;
d) allowing said heat
shocked cells to recover at a lower temperature than said heat shock temperature
and for a time sufficient to translate said transcribed structural gene,
producing said polypeptide or protein.
|
|
Claim 10
A method for increasing the production of a
structural polypeptide or protein in a transformed mammalian
cell comprising the steps of:
a) culturing a dihydrofolate reductase (DHFR) deficient mammalian
cell transformed with
(i) a DHFR gene under the control of a
constitutive promoter, and
(ii) a structural gene encoding for a
polypeptide or protein under the control of an inducible heat shock promoter, in
a culture medium containing methotrexate, for a time sufficient to permit growth
of said transformed mammalian cells;
b) inducing said heat shock promoter
at a temperature and for a time sufficient to transcribe said structural gene;
and
c) allowing said heat shocked cells to recover from said heat shock at
a lower temperature and for a time sufficient to translate said transcribed
structural gene, producing said polypeptide or protein.
|
|
Claim 17
A host cell cotransformed with:
a) a gene amplification system under the control of a constitutive promoter,
and
b) a structural gene encoding for a polypeptide or protein under
control of an inducible heat shock promoter.
|
Cold-regulated promoters
Patents filed by Takara Shuzo Co.
Actual granted independent claims
|
US
6479260
|
|
Claim 1
A vector which is characterized in containing
each of the following elements:
(1) a promoter which shows its action in the host to be used;
(2)
regulatory region for regulating the action of the promoter of (1); and
(3) a region which codes for the 5'-untranslated region derived from cold-shock
protein gene mRNA or a region which codes for the region where substitution,
deletion, insertion or addition of at least one base is applied to the said
untranslated region and
which contains a base sequence as shown in SEQ ID
NO:1 in the Sequence Listing.
|
|
Claim 10
An isolated promoter consisting of a base
sequence as shown in SEQ ID NO: 5 in the Sequence Listing.
|
|
Claim 11
An isolated promoter containing a base
sequence as shown in SEQ ID NO: 5 in the Sequence Listing and
consisting
of a base sequence having 135 or less bases,
wherein the promoter does not
contain the region which is ascribed to mRNA.
|
Actual filed independent claims
| EP 1033408 A1
& CA 2309600 AA
|
|
Claim 1
A vector which is characterized in containing
each of the following elements:
(1) a promoter which shows its action in the host to be used;
(2)
regulatory region for regulating the action of the promoter of (1); and
(3) a region which codes for the 5'-untranslated region derived from cold-shock
protein gene mRNA or a region which codes for the region where substitution,
deletion, insertion or addition of at least one base is applied to the said
untranslated region.
|
|
Claim 14
A promoter containing a base sequence as shown
in SEQ ID NO:5 in the Sequence Listing and
consisting of a base sequence
having 135 or less bases.
|
Patents filed by Japan Tobacco Inc.
Actual granted independent claims
|
US
6084089
|
|
Claim 1
A DNA sequence comprising a nucleotide sequence
from the first to the 3546th nucleotide in the nucleotide sequence shown in SEQ
ID NO:1.
|
|
Claim 2
A DNA sequence comprising a nucleotide sequence
from the 2418th to the 3541st nucleotide in the nucleotide sequence shown in SEQ
ID NO:1.
|
|
Claim 3
A DNA sequence comprising a nucleotide sequence
from the first to the 4120th nucleotide in the nucleotide sequence shown in SEQ
ID NO:2
|
|
AU
725727 B2*
|
|
Claim 1
A DNA sequence having a nucleotide sequence
from first to 3546th nucleotide in the nucleotide sequence shown in SEQ ID. No.
1, or a part thereof having a cold- inducible promoter activity, or a DNA
sequence having the same nucleotide sequence as said DNA sequences except that
one or more nucleotides are deleted or substituted, or one or more nucleotides
are inserted or added, which DNA sequence has a cold-inducible promoter
activity.
|
|
Claim 3
A DNA sequence having a nucleotide sequence
from 2418th to 3541st nucleotide in the nucleotide sequence shown in SEQ ID. No.
1, or a part thereof having a cold - inducible promoter activity, or a DNA
sequence having the same nucleotide sequence as the said DNA sequences except
that one or more nucleotides are deleted or substituted, or one or more
nucleotides are inserted or added, which DNA sequence has a cold-inducible
promoter activity.
|
|
Claim 4
A cold-inducible promoter sequence having a
nucleotide sequence from 2418th to 3541st nucleotide in the nucleotide sequence
shown in SEQ ID. No. 1, or a DNA sequence having the same nucleotide sequence as
said DNA sequence except that one or more nucleotides are deleted or
substituted, or one or more nucleotides are inserted 45 or added, which DNA
sequence has a cold-inducible promoter activity.
|
|
Claim 5
A DNA sequence having a nucleotide sequence
from first to 4 120th nucleotide in the nucleotide sequence shown in SEQ ID. No.
2, or a part thereof having a cold- inducible promoter activity, or a DNA
sequence having the same nucleotide sequence as the said DNA sequences except
that one or more nucleotides are del eted or substituted, or one or more
nucleotides are inserted or added, which DNA sequence has a cold-inducible
promoter activity.
|
|
Claim 7
A probe comprising a DNA fragment having at
least 18 consecutive nucleotides in the region from 45th to 839th nucleotide in
the sequence shown in SEQ ID N0:3 in the Sequence Listing or a sequence
complementary thereto.
|
|
*Independent claims as filed of the European
application EP 812917 A1 are the same as the granted claims of
the Australian patent. The only difference is in Claim 7 where instead of "
having at least 18 consecutive nucleotide", the probe recited
in the European application has "at least 15 consecutive
nucleotides".
|
Patents filed by Danisco
Actual granted independent claims
|
US
6184443
|
|
Claim 1
An isolated a-amylase promoter having
cold-sensitive promoter activity, having a sequence comprising the 5.5 Kb EcoRI
DNA fragment of Solanum tuberosum from the transformed E. coli
strain, DH5alpha-gPAmy 351 (NCIMB Accession Number 40682).
|
|
Claim 12
An isolated a-amylase promoter having
tuber-specific activity and having a sequence comprising SEQ ID NO: 1.
|
Actual filed independent claims
| EP 787194 A1
& CA 2202896 AA
|
|
Claim 1
A promoter comprising a nucleotide sequence
corresponding to the 5.5 Kb EcoR1 fragment isolated from Solanum
tuberosum or a variant, homologue or fragment thereof.
|
|
Claim 2
A promoter comprising a nucleotide sequence
corresponding to the 5.5 b EcoR1 fragment isolated from Solanum
tuberosum or a variant, homologue or fragment thereof, but wherein at least
a part of the promoter is inactivated.
|
|
Claim 3
A promoter comprising at least the nucleotide
sequence shown as Seq.I.D. No. 1 or a variant, homologue or fragment thereof.
|
|
Claim 4
A promoter comprising the nucleotide sequence
of any of one of the sequences shown as Seq.I.D.Nos. 4 - 17 or a variant,
homologue or fragment thereof.
|
|
Claim 5
A promoter comprising a nucleotide sequence
corresponding to the 5.5 Kb EcoR1 fragment isolated from Solanum
tuberosum or a variant, homologue or fragment thereof, but wherein at least
the nucleotide sequence shown as Seq.I.D. No. 1 is inactivated.
|
|
Claim 6
A promoter comprising a nucleotide sequence
corresponding to the 5.5 Kb EcoR1fragment isolated from Solanum
tuberosum or a variant, homologue or fragment thereof, but wherein at least
any of one of the sequences shown as Seq.I.D.Nos. 2-16 is inactivated.
|
Patents granted to the Canadian Ministry of Agriculture
Actual granted independent claims
|
US
5847102
|
|
Claim 1
An isolated DNA molecule comprising a 5'
regulatory region of a low temperature-responsive gene BN115 from Brassica
napus, comprising nucleotides 961-1210 of SEQ ID NO:1.
|
|
Claim 7
An isolated cold inducible promoter, or a
functional fragment thereof capable of regulating the expression of a gene in
response to a change in temperature,
said cold inducible promoter
comprising nucleotides 1-1271 of SEQ ID NO:1.
|
|
Claim 8
An isolated DNA molecule comprising a sequence
of at least 15 contiguous nucleotides of a 5' regulatory region of a low
temperature responsive gene BN115 from Brassica napus, as defined by
nucleotides 1-1271 of SEQ ID NO:1.
|
|
Claim 13
An isolated enhancer, or a functional fragment
thereof each capable of mediating the expression of a gene under the control of
a promoter and said enhancer or fragment,
said enhancer obtained from the
regulatory region of BN115 from Brassica napus.
|
|
Claim 16
An isolated negative regulatory element
capable of repressing gene expression at 22� C., obtained from the regulatory
region of BN115 from Brassica napus and comprising nucleotides 461-623
of SEQ ID NO:1.
|
| CA 2146712 |
|
Claim 1
An isolated DNA molecule comprising a 5'
regulatory region of a low temperature-responsive gene BN115 from Brassica
napus, comprising nucleotides 156-1362 of SEQ ID NO:1.
|
|
Claim 5
An isolated DNA molecule comprising a 5'
regulatory region of a low temperature-responsive gene BN115 from Brassica
napus, comprising nucleotides 1-1271 of SEQ ID NO:1.
|
|
Claim 7
An isolated cold inducible promoter, or a
functional fragment thereof capable of regulating the expression of a gene in
response to a change in temperature,
said cold inducible promoter
comprising nucleotides 1-1271 of SEQ ID NO:1.
|
|
Claim 8
An isolated DNA molecule comprising a sequence
defined by nucleotides 1-1271 of SEQ ID NO:1, a fragment thereof, or a
nucleotide sequence that hybridizes to said sequence defined by nucleotides
1-1271 of SEQ ID NO:1 under stringent hybridization conditions, wherein said DNA
molecule, said fragment thereof, or said nucleotide sequence exhibit low
temperature activity, said stringent hybridization conditions are selected from:
i) hybridization at 4X SSC at 42�C followed by washing in 0.1X SSC at 65�C
for an hour; and
ii) hybridization at 4X SSC, 50% formamide at 42�C
followed by washing in 0.1X SSC at 65�C for an hour.
|
|
Claim 13
An isolated enhancer comprising a sequence
defined by nucleotides 156-623 of SEQ ID NO:1, a functional fragment thereof, or
a nucleotide sequence that hybridizes to said sequence defined by nucleotides
156-623 of SEQ ID NO:1 under stringent hybridization conditions, wherein said
enhancer, fragment or nucleotide sequence are each capable of mediating
expression of a gene, said stringent hybridization conditions are selected from:
i) hybridization at 4X SSC at 42�C followed by washing in 0.1X SSC at 65�C
for an hour; and
ii) hybridization at 4X SSC, 50% formamide at 42�C
followed by washing in 0.1X SSC at 65�C for an hour.
|
|
Claim 16
An isolated negative regulatory element
capable of repressing gene expression at 22� C., obtained from the regulatory
region of BN115 from Brassica napus and comprising nucleotides 461-623
of SEQ ID NO:1.
|
Light-regulated promoters
A. Light-inducible
Patent granted to Calgene Inc.
|
US
5750385
|
|
Claim 1
A method for obtaining a plant having a
modified phenotype, said method comprising;
Transforming a host plant cell with a DNA construct under genomic integration
conditions, wherein said construct comprises as operably linked components in
the direction of transcription, a promoter region obtainable from a gene,
wherein transcription of said gene is light-inducible in a plant chloroplast
containing tissue, a DNA sequence of interest other than the native coding
sequence of said gene, and a transcription termination region, wherein said
components are functional in a plant cell,
Whereby said DNA construct becomes integrated into a genome of said plant
cell; regenerating a plant from said transformed plant cell, and growing said
plant under conditions whereby said DNA sequence of interest is expressed and a
plant having said modified phenotype is obtained.
|
|
Claim 2
A method for altering the phenotype of
chloroplast containing A DNA sequence comprising a light-inducible promoter
derived from a myxobacterium, and
a nucleotide sequence containing a
restriction site located to enable a nucleotide sequence which is to be
expressed to be placed under the control of the said promoter.
|
Patent granted to the University of Warwick
Actual granted independent claims
| CA 1321563 |
|
Claim 1
A recombinant DNA sequence containing a
light-inducible promoter derived from a myxobacterium.
|
|
Claim 2
A DNA sequence comprising a light-inducible
promoter derived from a myxobacterium, and
a nucleotide sequence
containing a restriction site located to enable a nucleotide sequence which is
to be expressed to be placed under the control of the said promoter.
|
|
Claim 8
A recombinant DNA sequence comprising a
light-inducible promoter derived from a myxobacterium,
operatively linked
to a DNA sequence coding for a polypeptide to be expressed,
the coding DNA
sequence not being operatively linked to the promoter in nature.
|
|
Claim 11
A method of producing a polypeptide which
comprises:
a) positioning a gene sequence coding for the polypeptide-under the op
erative control of a light-inducible promoter derived from a myxobacterium to
produce a recombinant DNA molecule;
b) transforming a competent bacterial
host with the said recombinant DNA molecule; and
c) subjecting the host to
light to induce the promoter whereby the gene is expressed.
|
B. Light-repressible
Patents filed by Suntory LTD
Actual granted independent claims
| NZ 508103 |
|
Claim 1
A DNA fragment containing the sequence of SEQ
ID NO: 1 as a core sequence,
whereby expression of a gene placed
downstream of said DNA fragment is repressed in the presence of light.
|
|
Claim 4
A promoter containing the nucleotide sequence
of SEQ ID NO: 1 as a core sequence,
whereby expression of a gene placed
downstream of said promoter is promoted in the dark but repressed in the
presence of light.
|
|
Note: Actual filed independent claims of patent applications
EP 1 077 257 & CA 2328139 AA are the same as the granted
independent claims of the New Zealander patent.
|
Patents granted to Mycogen Plant Science
Actual granted independent claims
|
US
5639952
|
|
Claim 1
A method for the expression of a structural
gene in a plant cell under conditions of darkness comprising the steps of:
a) transforming said plant cell with a recombinant molecule comprising:
a plant, dark- and light-active maize Cab promoter/regulatory system,
wherein said promoter/regulatory system is the promoter/regulatory system of Cab
AB1084 or cross-hybridizes with the promoter/regulatory system of Cab AB1 084
under stringent conditions, and
a heterologous plant-expressible structural
gene that is under the regulatory control of said plant Cab promoter/regulatory
system in said plant cell, and
b) maintaining conditions of darkness for
expression of said structural gene in said transformed plant cell.
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Claim 8
A method for enhancing the level of expression
obtained in the dark of a plant-expressible gene in a plant cell by exposing
said plant cell to conditions of illumination comprising the steps of:
a) transforming said plant cell with a recombinant molecule comprising:
a plant, dark- and light-active maize Cab promoter/regulatory system,
wherein said promoter/regulatory system is the promoter/regulatory system of Cab
AB1084 or cross-hybridizes with the promoter/regulatory system of Cab AB1084
under stringent conditions, and
a heterologous plant-expressible structural
gene that is under the regulatory control of said plant Cab promoter/regulatory
system in said plant cell, and
b) maintaining conditions of darkness for
expression of said structural gene in said transformed plant cell, and
c)
applying conditions of illumination to said transformed plant cell such that
enhancement by a factor of about 3- to 6-fold in expression of said structural
gene is obtained.
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Claim 15
A plant grown from a plant cell,
wherein said plant cell is transformed with a recombinant molecule comprising a
plant, dark- and light-active maize Cab promoter/regulatory system,
wherein said promoter/regulatory system is the promoter/regulatory system of Cab
AB1084 or cross-hybridizes with the promoter/regulatory system of Cab AB1084
under stringent conditions, and
a heterologous plant-expressible structural
gene that is under the regulatory control of said plant Cab promoter/regulatory
system in said plant cell.
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US
5656496
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Claim 1
A recombinant DNA molecule comprising:
a) a dark- and light-active Cab promoter/regulatory system which
cross-hybridizes with the Cab AB1084 promoter/regulatory system under stringent
conditions, and wherein said promoter/regulatory system:
(i) functions to direct the expression of a structural gene under its control
during conditions of darkness and
(ii) is stimulated to direct enhanced
expression of said structural gene under light conditions, and
b) a heterologous plant-expressible structural gene wherein said structural
gene is placed under the regulatory control of said plant promoter/regulatory
system.
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The information contained in this page was believed to be correct at the
time it was collated. New patents and patent applications, altered
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