Claims
- 1. An isolated nucleic acid comprising:
a Vibrio marinus nucleotide sequence selected from the group consisting of ORF 6 (SEQ ID NO:77), ORF 7 (SEQ ID NO:78), ORF 8 (SEQ ID NO:79), and ORF 9 (SEQ ID NO:80), as shown in FIG. 6.
- 2. An isolated nucleic acid comprising:
a nucleotide sequence which encodes a polypeptide of a polyketide-like synthesis system, wherein said system produces a docosahexenoic acid when expressed in a host cell.
- 3. The isolated nucleic acid according to claim 2, wherein said nucleotide sequence is derived from a marine bacterium.
- 4. An isolated nucleic acid according to claim 2, wherein said nucleotide sequence is derived from Schizochytrium.
- 5. The isolated nucleic acid according to claim 2, wherein said nucleotide sequence is a Vibrio marinus ORF 8 (SEQ ID NO:79), as shown in FIG. 6.
- 6. An isolated nucleic acid comprising a Schizochytrium nucleotide sequence comprising a sequence shown in a SEQ ID NO selected from the group consisting of SEQ ID NOS:69, 71 and 76.
- 7. An isolated nucleic acid comprising:
a nucleotide sequence which is substantially identical to a sequence of at least 50 nucleotides of a Vibrio marinus nucleotide sequence selected from the group consisting of ORF 6 (SEQ ID NO:77), ORF 7 (SEQ ID NO:78), ORF 8 (SEQ ID NO:79), and ORF 9 (SEQ ID NO:80), as shown in FIG. 6.
- 8. A recombinant microbial cell comprising at least one copy of an isolated nucleic acid according to claim 6.
- 9. The recombinant microbial cell according to claim 8, wherein said cell comprises each element of a polyketide-like synthesis system required to produce a long chain polyunsaturated fatty acid.
- 10. The recombinant microbial cell according to claim 9, wherein said cell is a eukaryotic cell.
- 11. The recombinant microbial cell according to claim 10, wherein said eukaryotic cell is a fungal cell, an algae cell or an animal cell.
- 12. The recombinant microbial cell according to claim 11, wherein said fungal cell is a yeast cell and said algae cell is a marine algae cell.
- 13. The recombinant microbial cell according to claim 8, wherein said cell is a prokaryotic cell.
- 14. The recombinant microbial cell according to claim 13, wherein said cell is a bacterial cell or a cyanobacterial cell.
- 15. A recombinant cell according to claim 14, wherein said bacterial cell is a lactobacillus cell.
- 16. The microbial cell according to claim 8, wherein said recombinant microbial cell is enriched for 22:6 fatty acids as compared to a non-recombinant microbial cell which is devoid of said isolated nucleic acid.
- 17. A method for production of docosahexenoic acid in a microbial cell culture, said method comprising:
growing a microbial cell culture having a plurality of microbial cells, wherein said microbial cells or ancestors of said microbial cells were transformed with a vector comprising one or more nucleic acids having a nucleotide sequence which encodes a polypeptide of a polyketide synthesizing system, wherein said one or more nucleic acids are operably linked to a promoter, under conditions whereby said one or more nucleic acids are expressed and docosahexenoic acid is produced in said microbial cell culture.
- 18. A method for production of a long chain polyunsaturated fatty acid in a plant cell, said method comprising:
growing a plant having a plurality of plant cells, wherein said plant cells or ancestors of said plant cells were transformed with a vector comprising one or more nucleic acids having a nucleotide sequence which encodes one or more polypeptides of a polyketide synthesizing system which produces a long chain polyunsaturated fatty acid, wherein each of said nucleic acids are operably linked to a promoter functional in a plant cell, under conditions whereby said polypeptides are expressed and a long chain polyunsaturated fatty acid is produced in said plant cells.
- 19. The method according to claim 17 or claim 18 wherein said nucleotide sequence is shown in a SEQ ID NO selected from the group consisting of SEQ ID NOS:69, 71 and 76.
- 20. The method according to claim 18, wherein said long chain polyunsaturated fatty acid produced in said plant cells is a 20:5 and 22:6 fatty acid.
- 21. The method according to claim 17, wherein said nucleotide sequence is selected from the group consisting of Vibrio marinus ORF 6 (SEQ ID NO:77), ORF 7 (SEQ ID NO:78), ORF 8 (SEQ ID NO:79), and ORF 9 (SEQ ID NO:80), as shown in FIG. 6 and Shewanella putrefaciens ORF 6 (SEQ ID NO:83), ORF 7 (SEQ ID NO:84), ORF 8 (SEQ ID NO:85), ORF 9 (SEQ ID NO:86), and ORF 3, which is complementary to SEQ ID NO:4, as shown in FIG. 4.
- 22. The method according to claim 18, wherein said nucleic acid constructs are derived from two or more polyketide synthesizing systems.
- 23. The method according to claim 18, wherein said long chain polyunsaturated fatty acid is eicosapentenoic acid.
- 24. The method according to claim 18, wherein said long chain polyunsaturated fatty acid is docosahexenoic acid.
- 25. A recombinant plant cell comprising:
one or more nucleic acids having a nucleotide sequence which encodes one or more polypeptides of a polyketide synthesizing system which produces a long chain polyunsaturated fatty acid, wherein each of said nucleic acids are operably linked to a promoter functional in said plant cell.
- 26. The recombinant plant cell according to claim 25, wherein said nucleotide sequence is shown in a SEQ ID NO selected from the group consisting of SEQ ID NOS: 69, 71 and 76.
- 27. The recombinant plant cell according to claim 26, wherein said recombinant plant cell is a recombinant seed cell.
- 28. The recombinant plant cell according to claim 27, wherein said recombinant seed cell is a recombinant embryo cell.
- 29. The recombinant plant cell according to claim 26, wherein said recombinant plant cell is from a plant selected from the group consisting of Brassica, soybean, safflower, and sunflower.
- 30. A plant oil produced by a recombinant plant cell according to claim 26.
- 31. The plant oil according to claim 30, wherein said plant oil comprises eicosapentenoic acid.
- 32. The plant oil according to claim 30, wherein said plant oil comprises docosahexenoic acid.
- 33. The plant oil according to claim 30, wherein said plant oil is encapsulated.
- 34. A dietary supplement comprising a plant oil according to claim 30.
- 35. A recombinant E. coli cell comprising:
one or more nucleic acids having a nucleotide sequence which encodes one or more polypeptides of a polyketide synthesizing system which produces a long chain polyunsaturated fatty acid, wherein each of said nucleic acids are operably linked to a promoter function in said E. coli cell.
- 36. The recombinant E. coli cell according to claim 35, wherein said long chain polyunsaturated fatty acid is docosahexenoic acid.
- 37. The recombinant E. coli cell according to claim 35, wherein said nucleotide sequence is shown in a SEQ ID NO selected from the group consisting of SEQ ID NOS: 69, 71 and 76.
- 38. A plant oil produced by a recombinant plant cell wherein said plant oil comprises a long chain polyunsaturated fatty acid exogenous to said plant oil, wherein said plant cell is produced according to a method comprising:
transforming said plant cell or an ancestor of said plant cell with a vector comprising one or more polypeptide of a polyketide synthesizing system which produces a long chain polyunsaturated fatty acid wherein each of said nucleic acids are operably linked to a promoter functional in said plant cell.
- 39. A plant oil according to claim 38, wherein said long chain polyunsaturated fatty acid is eicosapentenoic acid.
- 40. A plant oil according to claim 38, wherein said long chain polyunsaturated fatty acid is docosahexenoic acid.
CROSS-REFERENCE TO RELATED APPLICATION
[0001] This application is a continuation-in-part of U.S. Ser. No. 09/090,793, filed Jun. 4, 1998, which is a continuation-in-part of provisional application U.S. Ser. No. 60/048,650, filed Jun. 4, 1997, which is incorporated herein by reference.
Provisional Applications (1)
|
Number |
Date |
Country |
|
60048650 |
Jun 1997 |
US |
Continuations (1)
|
Number |
Date |
Country |
Parent |
09231899 |
Jan 1999 |
US |
Child |
10331061 |
Dec 2002 |
US |
Continuation in Parts (1)
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Number |
Date |
Country |
Parent |
09090793 |
Jun 1998 |
US |
Child |
09231899 |
Jan 1999 |
US |