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.
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Patent No
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Title, Independent Claims and Summary
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Applicant
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AU
2001/32896
- Filed - 19 January 2001
- Granted - under examination
- Lapsed on 31 Aug 2006
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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).
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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).
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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).
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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).
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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).
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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.
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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.
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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.
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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).
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Pioneer Hi-Bred
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Remarks
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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.
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