The University of Georgia Research Foundation patent family
University of Georgia Research Foundation, Inc. has a granted
patent in the United States on methods and materials for selecting transgenic
cells based on arabitol or ribitol as positive selectable markers. Patent
application was also filed in Australia.[add a comment]
The invention relates to isolated polynucleotide molecules coding
for a protein possessing arabitol/ribitol dehydrogenase enzymatic activity and a
protein possessing arabitol/ribitol kinase enzymatic activity. It also relates
to a positive selection system that involves conferring to transformed cells
(includes protoplasts, as well as cells of plants, animals and microorganisms)
the ability to metabolize arabitol, ribitol and/or mannitol and selecting the
transformed cells.[add a comment]
Technology overview
D-arabitol and ribitol are two of the four possible pentitols (five-carbon
sugar alcohol, C5H12O5) formed by the reduction
of ribose. In 2001, LaFayette and Parrott reported that E. coli strains B and
K-12 cannot metabolize these pentitols. However, E. coli strain C originally
isolated in the Lister Institute, London, in 1920 (NCTC 1983) can metabolize
both D-arabitol and ribitol and thus grow on these pentitols when they are the
sole carbon source. This ability is due to the presence of two genes coding for
arabitol deydrogenase and ribitol dehydrogenase that convert arabitol and
ribitol into xylulose and ribulose, respectively.
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| The structure of a pentitol |
Plants are not able to metabolize arabitol and ribitol. The positive
selection strategy in this aspect is to engineer plant cells by introducing an
arabitol or ribitol dehydrogenase gene so that the transformed cells can utilise
arabitol or ribitol as carbon source. For example, when a gene coding for a
bacterial arabitol dehydrogenase is transferred into and expressed in a plant
cell, the cell will be able to grow in a medium containing D-arabitol by
converting arabitol into the plant-metabolizable xylulose, whereas an
untransformed plant cell will not proliferate.
Specific patent information
| Patent/Application Number |
Title, Independent Claims and Summary |
Assignee |
|
US
7005561 B2
- Earliest priority - 8 Mar 2000
- Filed - 3 Mar 2001
- Granted - 28 Feb 2006
- Expected expiry - 8 Aug 2021 (see disclaimer in the patent)
|
Title - Arabitol or ribitol as positive selectable markers
Claim 1
An isolated polynucleotide molecule comprising at least one gene of interest
and at least one selectable marker gene, wherein said at least one selectable
marker gene comprises a nucleotide sequence which selectively hybridizes under
high stringency conditions to the complement of a nucleotide sequence shown in
SEQ
ID NO: 2, or a plant optimized version thereof, wherein said nucleotide
sequence encodes for a protein possessing ribitol dehydrogenase enzymatic
activity and a protein possessing ribitol kinase enzymatic activity.
|
Claim 6
A method of selecting transformed cells from a population of cells
comprising;
a) introducing into the genome of a cell a gene of interest and a selectable
marker gene;
b) obtaining transformed cells;
c) supplying to the population of cells a marker compound wherein said
transformed cells have a selective advantage over non-transformed cells due to
expression or transcription of the the selectable marker gene in the presence of
the marker compound; and
d) selecting said transformed cells from the population of cells; wherein said
selectable marker gene comprises a nucleotide sequence which selectively
hybridizes under high stringency conditions to the complement of a nucleotide
sequence shown in
SEQ
ID NO: 2, or a plant optimized version thereof, wherein said nucleotide
sequence encodes a protein that possesses ribitol dehydrogenase enzymatic
activity and a protein that possesses ribitol kinase enzymatic activity; and
said marker compound comprises arabitol, ribitol, or mannitol.
|
|
Claim 12
An isolated polynucleotide molecule comprising a nucleotide sequence which
selectively hybridizes under high stringency conditions to the complement of a
plant optimized version of the nucleotide sequences shown in
SEQ
ID NO: 2, and wherein said nucleotide sequence encodes for a protein
possessing ribitol dehydrogenase activity and a protein possessing ribitol
kinase activity.
|
|
Claim 13
An isolated polynucleotide molecule comprising at least one gene of interest,
and at least one selectable marker gene, wherein said at least one selectable
marker gene comprises a nucleotide sequence encoding
SEQ
ID NOS.: 3 and
4.
|
|
Claim 14
An isolated polynucleotide molecule comprising at least one gene of interest,
and at least one selectable marker gene, wherein said at least one selectable
marker gene comprises a nucleotide sequence which selectively hybridizes under
high stringency conditions to the complement of a nucleotide sequence shown in
SEQ
ID NO: 1, or a plant optimized version thereof, wherein said at least one
selectable marker gene encodes for a protein possessing arabitol dehydrogenase
enzymatic activity.
|
|
Claim 15
A method of selecting transformed cells from a population of cells comprising
a) introducing into the genome of a cell a gene of interest and a selectable
marker gene;
b) obtaining transformed cells;
c) supplying to the population of cells a marker compound wherein said
transformed cells have a selective advantage over non-transformed cells due to
expression or transcription of the selectable marker gene in the presence of the
marker compound; and
d) selecting said transformed cells from the population of cells; wherein said
selectable marker gene comprises a nucleotide sequence which selectively
hybridizes under high stringency conditions to the complement of a nucleotide
sequence shown in
SEQ
ID NO: 1, or a plant optimized version thereof, and encodes a protein having
arabitol dehydrogenase enzymatic activity; and wherein said marker compound is
arabitol.
|
|
Claim 16
A method of selecting transformed cells from a population of cells comprising
a) introducing into the genome of a cell a gene of interest and a selectable
marker gene;
b) obtaining transformed cells;
c) supplying to the population of cells a marker compound wherein said
transformed cells have a selective advantage over non-transformed cells due to
expression or transcription of the selectable marker gene in the presence of the
marker compound; and
d) selecting said transformed cells from the population of cells; wherein said
selectable marker gene comprises a nucleotide sequence encoding
SEQ
ID NO.: 3, and a nucleotide sequence encoding
SEQ
ID NO.: 4; and wherein said marker compound is ribitol.
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University of Georgia Research Foundation
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Remarks
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The related patent application in Australia (AU 200140117)
has lapsed. A PCT application was also filed
(WO
2001/66779).
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Search strategy
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Search details
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Date of search
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31/05/2006
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Database searched
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Patent Lens
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Type of search
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Simple, stemming on
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Collections searched
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AU-B, US-A, US-B, EP-B, WO
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Search terms
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"Positive AND selection AND transformation AND cell" in abstract
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Results
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117 hits
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Comments
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This search results in patents from several patent families that related to
the positive selection topic:
1. the Syngenta family represented by WO 1993/05163
2. the "University of Georgia Research Foundation, Inc." family represented
by US 7005561
titled "Arabitol or ribitol as positive selectable markers".
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