CA2416602A1 - Nucleotide sequences of a new class of diverged delta-9 stearoyl-acp desaturase genes - Google Patents
Nucleotide sequences of a new class of diverged delta-9 stearoyl-acp desaturase genes Download PDFInfo
- Publication number
- CA2416602A1 CA2416602A1 CA002416602A CA2416602A CA2416602A1 CA 2416602 A1 CA2416602 A1 CA 2416602A1 CA 002416602 A CA002416602 A CA 002416602A CA 2416602 A CA2416602 A CA 2416602A CA 2416602 A1 CA2416602 A1 CA 2416602A1
- Authority
- CA
- Canada
- Prior art keywords
- seq
- delta
- fatty acid
- polypeptide
- isolated polynucleotide
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
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Classifications
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/0004—Oxidoreductases (1.)
- C12N9/0071—Oxidoreductases (1.) acting on paired donors with incorporation of molecular oxygen (1.14)
- C12N9/0083—Miscellaneous (1.14.99)
Abstract
An isolated nucleic acid fragment encoding a diverged delta-9 fatty acid desaturase is disclosed. Also the construction of a chimeric gene encoding a ll or a portion of the diverged delta-9 fatty acid desaturase is disclosed, in sense or antisense orientation, wherein expression of the chimeric gene results in production of altered levels of the diverged delta-9 fatty acid desaturase in a transformed host cell.
Claims
What is claimed is:
1. An isolated polynucleotide comprising:
(a) a nucleotide sequence encoding a polypeptide having delta-9 fatty acid desaturase activity that has at least 80% identity based on the Clustal method of alignment when compared to a polypeptide selected from the group consisting of SEQ ID NO:2, 4, 6, 8, 10, 12, 14, or 16; or (b) the complement of (a).
2. An isolated polynucleotide comprising:
(a) a nucleotide sequence encoding a polypeptide having delta-9 fatty acid desaturase activity that has at least 85% identity based on the Clustal method of alignment when compared to a polypeptide selected from the group consisting of SEQ ID NO:2, 4, 6, 8, 10, 12, 14, or 16; or (b) the complement of (a).
3. An isolated polynucleotide comprising:
(a) a nucleotide sequence encoding a polypeptide having delta-9 fatty acid desaturase activity that has at least 90% identity based on the Clustal method of alignment when compared to a polypeptide selected from the group consisting of SEQ ID NO:2, 4, 6, 8, 10, 12, 14, or 16; or (b) the complement of (a).
4. An isolated polynucleotide comprising:
(a) a nucleotide sequence encoding a polypeptide having delta-9 fatty acid desaturase activity that has at least 95% identity based on the Clustal method of alignment when compared to a polypeptide selected from the group consisting of SEQ ID NO:2, 4, 6, 8, 10, 12, 14, or 16; or (b) the complement of (a).
5. The isolated polynucleotide of any of Claims 1-4, wherein the nucleotide sequence comprises a nucleic acid sequence selected from the group consisting of SEQ ID NO:1,3,5,7,9,11,13,or15.
6. A chimeric construct comprising the isolated polynucleotide of any of Claims 1-4 operably linked to at least one suitable regulatory sequence.
7. A chimeric construct comprising the isolated polynucleotide of Claim 5 operably linked to at least one suitable regulatory sequence.
8. A host cell comprising the chimeric construct of Claim 6.
9. A host cell comprising the chimeric construct of Claim 7.
10. A method of obtaining a nucleic acid fragment encoding a delta-9 fatty acid desaturase polypeptide comprising the steps of:
(a) probing a cDNA or genomic library with an isolated polynucleotide comprising at least one of 30 contiguous nucleotides derived from a nucleotide sequence selected from the group consisting of SEQ ID
NO:1, 3, 5, 7, 9, 11, 13, or 15 and/or a complement of the nucleotide sequence;
(b) identifying a DNA clone that hybridizes with the isolated polynucleotide;
(c) isolating the identified DNA clone; and (d) sequencing a cDNA or genomic fragment that comprises the isolated DNA clone.
11. A method of obtaining a nucleic acid fragment encoding a delta-9 fatty acid desaturase polypeptide comprising the steps of:
(a) probing a cDNA or genomic library with an isolated polynucleotide comprising at least one of 30 contiguous nucleotides derived from a nucleotide sequence selected from the group consisting of SEQ ID
NO:1, 3, 5, 7, 9, 11, 13, or 15 and/or a complement of such nucleotide sequences;
(b) identifying a DNA clone that hybridizes with the isolated polynucleotide;
(c) isolating the identified DNA clone;
(d) inserting the DNA clone into a plasmid suitable for expression in a bacterial or yeast cell; and (e) assaying for delta-9 desaturase activity or alterations in fatty acid composition of the host cell.
12. A method of identifying an isolated polynucleotide that encodes a delta-9 fatty acid desaturase comprising the steps of:
(a) determining an amino acid sequence of the polypeptide encoded by the isolated DNA;
(b) determining if the amino acid sequence comprises at least two amino acid sequences selected from the group consisting of HSMPPEK corresponding to amino acids 67-73 of SEQ ID NO:2, LPLLKPVE corresponding to amino acids 89-96 of SEQ ID NO:2, EYFVVLVGDM corresponding to amino acids 132-141 of SEQ ID
NO:2, EKTV corresponding to amino acids 205-208 of SEQ ID
NO:2, GMDPGT corresponding to amino acids 215-220 of SEQ ID
NO:2, NNPYLGFVYTSFQERAT corresponding to amino acids 222-238 of SEQ ID NO:2, VLAR corresponding to amino acids 256-259 of SEQ ID NO:2, RIVE corresponding to amino acids 277-280 of SEQ ID NO:2, ITMPAHL corresponding to amino acids 302-308 of SEQ ID NO:2, or DFVCGLA corresponding to amino acids 364-370 of SEQ ID NO:2.
13. A method of identifying an isolated polynucleotide that encodes a delta-9 fatty acid desaturase comprising the steps of:
(a) determining the polypeptide sequence of Claim 10, 11, or 12;
(b) determining that the amino acid sequence of the polypeptide does not contain at least one of the following amino acid sequences KEIPDDYFWLVGDMITEEALPTYQTMLNT corresponding to positions 116-145 of SEQ ID NO:23; or DYADILEFLVGRWK
corresponding to positions 324-337 of SEQ ID NO:23.
14. A method of altering the level of expression of a delta-9 fatty acid desaturase in a host cell comprising:
(a) transforming a host cell with the chimeric gene of Claim 6; and (b) growing the transformed host cell produced in step (a) under conditions that are suitable for expression of the chimeric construct wherein expression of the chimeric construct results in production of altered levels of a delta-9 fatty acid desaturase in the transformed host cell.
15. A method of altering the level of expression of a delta-9 fatty acid desaturase in a host cell comprising:
(a) transforming a host cell with the chimeric construct of Claim 6 ; and (b) growing the transformed host cell produced in step (a) under conditions that are suitable for expression of the chimeric construct wherein expression of the chimeric gene results in production of altered levels of a delta-9 fatty acid desaturase in the transformed host cell.
1. An isolated polynucleotide comprising:
(a) a nucleotide sequence encoding a polypeptide having delta-9 fatty acid desaturase activity that has at least 80% identity based on the Clustal method of alignment when compared to a polypeptide selected from the group consisting of SEQ ID NO:2, 4, 6, 8, 10, 12, 14, or 16; or (b) the complement of (a).
2. An isolated polynucleotide comprising:
(a) a nucleotide sequence encoding a polypeptide having delta-9 fatty acid desaturase activity that has at least 85% identity based on the Clustal method of alignment when compared to a polypeptide selected from the group consisting of SEQ ID NO:2, 4, 6, 8, 10, 12, 14, or 16; or (b) the complement of (a).
3. An isolated polynucleotide comprising:
(a) a nucleotide sequence encoding a polypeptide having delta-9 fatty acid desaturase activity that has at least 90% identity based on the Clustal method of alignment when compared to a polypeptide selected from the group consisting of SEQ ID NO:2, 4, 6, 8, 10, 12, 14, or 16; or (b) the complement of (a).
4. An isolated polynucleotide comprising:
(a) a nucleotide sequence encoding a polypeptide having delta-9 fatty acid desaturase activity that has at least 95% identity based on the Clustal method of alignment when compared to a polypeptide selected from the group consisting of SEQ ID NO:2, 4, 6, 8, 10, 12, 14, or 16; or (b) the complement of (a).
5. The isolated polynucleotide of any of Claims 1-4, wherein the nucleotide sequence comprises a nucleic acid sequence selected from the group consisting of SEQ ID NO:1,3,5,7,9,11,13,or15.
6. A chimeric construct comprising the isolated polynucleotide of any of Claims 1-4 operably linked to at least one suitable regulatory sequence.
7. A chimeric construct comprising the isolated polynucleotide of Claim 5 operably linked to at least one suitable regulatory sequence.
8. A host cell comprising the chimeric construct of Claim 6.
9. A host cell comprising the chimeric construct of Claim 7.
10. A method of obtaining a nucleic acid fragment encoding a delta-9 fatty acid desaturase polypeptide comprising the steps of:
(a) probing a cDNA or genomic library with an isolated polynucleotide comprising at least one of 30 contiguous nucleotides derived from a nucleotide sequence selected from the group consisting of SEQ ID
NO:1, 3, 5, 7, 9, 11, 13, or 15 and/or a complement of the nucleotide sequence;
(b) identifying a DNA clone that hybridizes with the isolated polynucleotide;
(c) isolating the identified DNA clone; and (d) sequencing a cDNA or genomic fragment that comprises the isolated DNA clone.
11. A method of obtaining a nucleic acid fragment encoding a delta-9 fatty acid desaturase polypeptide comprising the steps of:
(a) probing a cDNA or genomic library with an isolated polynucleotide comprising at least one of 30 contiguous nucleotides derived from a nucleotide sequence selected from the group consisting of SEQ ID
NO:1, 3, 5, 7, 9, 11, 13, or 15 and/or a complement of such nucleotide sequences;
(b) identifying a DNA clone that hybridizes with the isolated polynucleotide;
(c) isolating the identified DNA clone;
(d) inserting the DNA clone into a plasmid suitable for expression in a bacterial or yeast cell; and (e) assaying for delta-9 desaturase activity or alterations in fatty acid composition of the host cell.
12. A method of identifying an isolated polynucleotide that encodes a delta-9 fatty acid desaturase comprising the steps of:
(a) determining an amino acid sequence of the polypeptide encoded by the isolated DNA;
(b) determining if the amino acid sequence comprises at least two amino acid sequences selected from the group consisting of HSMPPEK corresponding to amino acids 67-73 of SEQ ID NO:2, LPLLKPVE corresponding to amino acids 89-96 of SEQ ID NO:2, EYFVVLVGDM corresponding to amino acids 132-141 of SEQ ID
NO:2, EKTV corresponding to amino acids 205-208 of SEQ ID
NO:2, GMDPGT corresponding to amino acids 215-220 of SEQ ID
NO:2, NNPYLGFVYTSFQERAT corresponding to amino acids 222-238 of SEQ ID NO:2, VLAR corresponding to amino acids 256-259 of SEQ ID NO:2, RIVE corresponding to amino acids 277-280 of SEQ ID NO:2, ITMPAHL corresponding to amino acids 302-308 of SEQ ID NO:2, or DFVCGLA corresponding to amino acids 364-370 of SEQ ID NO:2.
13. A method of identifying an isolated polynucleotide that encodes a delta-9 fatty acid desaturase comprising the steps of:
(a) determining the polypeptide sequence of Claim 10, 11, or 12;
(b) determining that the amino acid sequence of the polypeptide does not contain at least one of the following amino acid sequences KEIPDDYFWLVGDMITEEALPTYQTMLNT corresponding to positions 116-145 of SEQ ID NO:23; or DYADILEFLVGRWK
corresponding to positions 324-337 of SEQ ID NO:23.
14. A method of altering the level of expression of a delta-9 fatty acid desaturase in a host cell comprising:
(a) transforming a host cell with the chimeric gene of Claim 6; and (b) growing the transformed host cell produced in step (a) under conditions that are suitable for expression of the chimeric construct wherein expression of the chimeric construct results in production of altered levels of a delta-9 fatty acid desaturase in the transformed host cell.
15. A method of altering the level of expression of a delta-9 fatty acid desaturase in a host cell comprising:
(a) transforming a host cell with the chimeric construct of Claim 6 ; and (b) growing the transformed host cell produced in step (a) under conditions that are suitable for expression of the chimeric construct wherein expression of the chimeric gene results in production of altered levels of a delta-9 fatty acid desaturase in the transformed host cell.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CA2708605A CA2708605C (en) | 2000-08-22 | 2001-08-22 | Nucleotide sequences of a new class of diverged delta-9 stearoyl-acp desaturase genes |
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US22699600P | 2000-08-22 | 2000-08-22 | |
| US60/226,996 | 2000-08-22 | ||
| PCT/US2001/026246 WO2002016565A2 (en) | 2000-08-22 | 2001-08-22 | Nucleotide sequences of a new class of diverged delta-9 stearoyl-acp desaturase genes |
Related Child Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA2708605A Division CA2708605C (en) | 2000-08-22 | 2001-08-22 | Nucleotide sequences of a new class of diverged delta-9 stearoyl-acp desaturase genes |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CA2416602A1 true CA2416602A1 (en) | 2002-02-28 |
| CA2416602C CA2416602C (en) | 2010-10-26 |
Family
ID=22851321
Family Applications (2)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA2708605A Expired - Fee Related CA2708605C (en) | 2000-08-22 | 2001-08-22 | Nucleotide sequences of a new class of diverged delta-9 stearoyl-acp desaturase genes |
| CA2416602A Expired - Fee Related CA2416602C (en) | 2000-08-22 | 2001-08-22 | Nucleotide sequences of a new class of diverged delta-9 stearoyl-acp desaturase genes |
Family Applications Before (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA2708605A Expired - Fee Related CA2708605C (en) | 2000-08-22 | 2001-08-22 | Nucleotide sequences of a new class of diverged delta-9 stearoyl-acp desaturase genes |
Country Status (6)
| Country | Link |
|---|---|
| US (3) | US20030054521A1 (en) |
| EP (1) | EP1311659B1 (en) |
| AU (3) | AU2001286627A1 (en) |
| BR (1) | BR0113563A (en) |
| CA (2) | CA2708605C (en) |
| WO (1) | WO2002016565A2 (en) |
Families Citing this family (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6511392B1 (en) * | 1999-02-08 | 2003-01-28 | Haroon Chohan | Baseball bat with interchangeable portions |
| DE60144517D1 (en) * | 2000-06-23 | 2011-06-09 | Pioneer Hi Bred Int | RECOMBINANT CONSTRUCTS AND THEIR USE IN REDUCING GENE EXPRESSION |
| CA2450000A1 (en) | 2003-12-18 | 2005-06-18 | Alberta Research Council Inc. | Method of creating plants with reduced level of saturated fatty acid in seed oil |
| BR112013000739A2 (en) | 2010-07-14 | 2016-06-07 | E I Pont De Nemours And Company | "soy protein product, first soy protein product, food, beverage, animal feed, method for improving the drying efficiency of a soy protein product and method of preparing a soy protein product" |
| US9598701B2 (en) | 2012-01-23 | 2017-03-21 | E I Du Pont De Nemours And Company | Down-regulation of gene expression using artificial MicroRNAs for silencing fatty acid biosynthetic genes |
Family Cites Families (16)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5190931A (en) * | 1983-10-20 | 1993-03-02 | The Research Foundation Of State University Of New York | Regulation of gene expression by employing translational inhibition of MRNA utilizing interfering complementary MRNA |
| US4945050A (en) * | 1984-11-13 | 1990-07-31 | Cornell Research Foundation, Inc. | Method for transporting substances into living cells and tissues and apparatus therefor |
| US4683195A (en) * | 1986-01-30 | 1987-07-28 | Cetus Corporation | Process for amplifying, detecting, and/or-cloning nucleic acid sequences |
| US5283323A (en) * | 1985-08-07 | 1994-02-01 | The United States Of America As Represented By The Department Of Health And Human Services | Method of producing immune response |
| US4800159A (en) * | 1986-02-07 | 1989-01-24 | Cetus Corporation | Process for amplifying, detecting, and/or cloning nucleic acid sequences |
| ES2018274T5 (en) | 1986-03-11 | 1996-12-16 | Plant Genetic Systems Nv | VEGETABLE CELLS RESISTANT TO GLUTAMINE SYNTHETASE INHIBITORS, PREPARED BY GENETIC ENGINEERING. |
| US5107065A (en) * | 1986-03-28 | 1992-04-21 | Calgene, Inc. | Anti-sense regulation of gene expression in plant cells |
| US5015580A (en) | 1987-07-29 | 1991-05-14 | Agracetus | Particle-mediated transformation of soybean plants and lines |
| US5231020A (en) * | 1989-03-30 | 1993-07-27 | Dna Plant Technology Corporation | Genetic engineering of novel plant phenotypes |
| WO1991013972A1 (en) | 1990-03-16 | 1991-09-19 | Calgene, Inc. | Plant desaturases - compositions and uses |
| US7037692B1 (en) * | 1990-03-16 | 2006-05-02 | Calgene, Inc. | Plant desaturases compositions and uses |
| AU659087B2 (en) | 1990-05-25 | 1995-05-11 | E.I. Du Pont De Nemours And Company | Nucleotide sequence of soybean stearoyl-ACP desaturase gene |
| DE69233118T2 (en) | 1991-12-04 | 2004-04-15 | E.I. Du Pont De Nemours And Co., Wilmington | FATTY ACID DESATURASE GENES FROM PLANTS |
| EP0668919B1 (en) | 1992-11-17 | 2003-06-04 | E.I. Du Pont De Nemours And Company | Genes for microsomal delta-12 fatty acid desaturases and related enzymes from plants |
| US5760206A (en) * | 1992-12-11 | 1998-06-02 | E. I. Du Pont De Nemours And Company | Nucleotide sequence of soybean stearoyl-ACP desaturase gene |
| DE60144517D1 (en) * | 2000-06-23 | 2011-06-09 | Pioneer Hi Bred Int | RECOMBINANT CONSTRUCTS AND THEIR USE IN REDUCING GENE EXPRESSION |
-
2001
- 2001-08-22 CA CA2708605A patent/CA2708605C/en not_active Expired - Fee Related
- 2001-08-22 CA CA2416602A patent/CA2416602C/en not_active Expired - Fee Related
- 2001-08-22 AU AU2001286627A patent/AU2001286627A1/en not_active Abandoned
- 2001-08-22 US US09/934,900 patent/US20030054521A1/en not_active Abandoned
- 2001-08-22 EP EP01966086A patent/EP1311659B1/en not_active Expired - Lifetime
- 2001-08-22 BR BR0113563-5A patent/BR0113563A/en not_active IP Right Cessation
- 2001-08-22 WO PCT/US2001/026246 patent/WO2002016565A2/en active IP Right Grant
-
2004
- 2004-11-04 US US10/981,293 patent/US7498427B2/en not_active Expired - Fee Related
-
2007
- 2007-09-19 AU AU2007216843A patent/AU2007216843B2/en not_active Ceased
-
2009
- 2009-01-22 US US12/357,474 patent/US20090203142A1/en not_active Abandoned
- 2009-06-03 AU AU2009202217A patent/AU2009202217A1/en not_active Withdrawn
Also Published As
| Publication number | Publication date |
|---|---|
| BR0113563A (en) | 2004-02-03 |
| EP1311659B1 (en) | 2006-03-08 |
| AU2009202217A1 (en) | 2009-06-25 |
| US20090203142A1 (en) | 2009-08-13 |
| WO2002016565A2 (en) | 2002-02-28 |
| WO2002016565A3 (en) | 2002-12-12 |
| CA2416602C (en) | 2010-10-26 |
| US20050066390A1 (en) | 2005-03-24 |
| EP1311659A2 (en) | 2003-05-21 |
| CA2708605A1 (en) | 2002-02-28 |
| US20030054521A1 (en) | 2003-03-20 |
| CA2708605C (en) | 2013-04-23 |
| AU2007216843B2 (en) | 2009-08-13 |
| US7498427B2 (en) | 2009-03-03 |
| AU2007216843A1 (en) | 2007-10-11 |
| AU2001286627A1 (en) | 2002-03-04 |
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Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| EEER | Examination request | ||
| MKLA | Lapsed |
Effective date: 20160822 |