Claims
- 1. An isolated DNA sequence encoding an enzyme active in the formation of triacylglycerol from diacylglycerol and fatty acyl substrates.
- 2. The isolated DNA sequence according to claim 1, wherein said nucleic acid sequence encodes diacylglycerol acyltransferase.
- 3. The isolated DNA sequence according to claim 1, wherein said nucleic acid sequence is isolated from a eukaryotic cell source.
- 4. The isolated DNA sequence according to claim 3, wherein said eukaryotic cell source is selected from the group consisting of mammalian, nematode, fungal, and plant cells.
- 5. The DNA encoding sequence of claim 4 wherein said diacylglycerol acyltransferase protein is from Mortierella ramanniana.
- 6. The DNA encoding sequence of claim 4 wherein said cell source is selected from the group consisting of soybean, Brassica napus, Arabidopsis thaliana, corn, human, murine, Mortierella alpina, Aspergillus fumigatus, Aspergillus oraceus, Fuserium graminearum, Schzichytrium aggregatum, Caenorhabditis elegans, and Sacchromyces cervaisae.
- 7. The DNA encoding sequence of claim 4 wherein said enzyme is selective for synthesis of a structured triacylglycerol.
- 8. The DNA encoding sequence of claim 7 wherein said structured triacylglycerol has a structure of the formula: S-U-S, wherein S represents a saturated fatty acid and U represents an unsaturated fatty acid.
- 9. The DNA encoding sequence of claim 4 wherein said diacylglycerol acyltransferase protein is encoded by a sequence which includes a nucleotide sequence selected from the group consisting of SEQ ID Nos: 46 through 72, inclusive.
- 10. An isolated polypeptide comprising the amino acid sequence of SEQ ID NO: 38.
- 11. An isolated polypeptide comprising the amino acid sequence of SEQ ID NO: 45.
- 12. An isolated polypeptide encoded by the nucleotide sequence as set forth in SEQ ID NO:37.
- 13. An isolated polypeptide encoded by the nucleotide sequence as set forth in SEQ ID NO:44.
- 14. An isolated polypeptide encoded by the nucleotide sequence as set forth in SEQ ID NO:84.
- 15. An isolated polynucleotide selected from the group consisting of:
a) an isolated polynucleotide comprising a nucleotide sequence encoding the polypeptide of SEQ ID NO: 38; b) an isolated polynucleotide comprising SEQ ID NO: 37; c) an isolated polynucleotide comprising a nucleotide sequence which has at least 70% identity to that of SEQ ID NO: 37 over the entire length of SEQ ID NO: 37; d) an isolated polynucleotide comprising a nucleotide sequence which has at least 80% identity to that of SEQ ID NO: 37 over the entire length of SEQ ID NO: 37; e) an isolated polynucleotide comprising a nucleotide sequence which has at least 90% identity to that of SEQ ID NO: 37 over the entire length of SEQ ID NO: 37; f ) an isolated polynucleotide comprising a nucleotide sequence which has at least 95% identity to that of SEQ ID NO: 37 over the entire length of SEQ ID NO: 37; g) an isolated polynucleotide that hybridizes, under stringent conditions, to SEQ ID NO: 37 or a fragment thereof; and h) an isolated polynucleotide complementary to the polynucleotide sequence of (a), (b), (c), (d), (e), (f), or (g).
- 16. An isolated polynucleotide selected from the group consisting of:
a) an isolated polynucleotide comprising SEQ ID NO: 84; b) an isolated polynucleotide comprising a nucleotide sequence which has at least 70% identity to that of SEQ ID NO: 84 over the entire length of SEQ ID NO: 84; c) an isolated polynucleotide comprising a nucleotide sequence which has at least 80% identity to that of SEQ ID NO: 84 over the entire length of SEQ ID NO: 84; d) an isolated polynucleotide comprising a nucleotide sequence which has at least 90% identity to that of SEQ ID NO: 84 over the entire length of SEQ ID NO: 84; e) an isolated polynucleotide comprising a nucleotide sequence which has at least 95% identity to that of SEQ ID NO: 84 over the entire length of SEQ ID NO: 84; f) an isolated polynucleotide that hybridizes, under stringent conditions, to SEQ ID NO: 84 or a fragment thereof; and g) an isolated polynucleotide complementary to the polynucleotide sequence of (a), (b), (c), (d), (e), or (f).
- 17. An isolated polynucleotide selected from the group consisting of:
a) an isolated polynucleotide comprising a nucleotide sequence encoding the polypeptide of SEQ ID NO: 45; b) an isolated polynucleotide comprising SEQ ID NO: 44; c) an isolated polynucleotide comprising a nucleotide sequence which has at least 70% identity to that of SEQ ID NO: 44 over the entire length of SEQ ID NO: 44; d) an isolated polynucleotide comprising a nucleotide sequence which has at least 80% identity to that of SEQ ID NO: 44 over the entire length of SEQ ID NO: 44; e) an isolated polynucleotide comprising a nucleotide sequence which has at least 90% identity to that of SEQ ID NO: 44 over the entire length of SEQ ID NO: 44; f ) an isolated polynucleotide comprising a nucleotide sequence which has at least 95% identity to that of SEQ ID NO: 44 over the entire length of SEQ ID NO: 44; g) an isolated polynucleotide that hybridizes, under stringent conditions, to SEQ ID NO: 44 or a fragment thereof; and h) an isolated polynucleotide complementary to the polynucleotide sequence of (a), (b), (c), (d), (e), (f) or (g).
- 18. An isolated polynucleotide selected from the group consisting of SEQ ID Nos: 46 through 67, inclusive.
- 19. A Mortierella acyltransferase protein wherein said acyltransferase is active in the formation of triacylglycerol from diacylglycerol and fatty acyl substrates and wherein said acyltransferase is isolated from other proteins indigenous to Mortierella.
- 20. An acyltransferase protein, wherein said protein has an apparent molecular mass of about 36 kD on SDS-PAGE, said protein being substantially purified away from membranes and other proteins of the native cell and capable of catalyzing the production of triglycerides from diacylglycerol and fatty acyl substrates.
- 21. An acyltransferase protein, wherein said protein has an apparent molecular mass of about 36.5 kD on SDS-PAGE, said protein being substantially purified away from membranes and other proteins of the native cell and capable of catalyzing the production of triglycerides from diacylglycerol and fatty acyl substrates.
- 22. A nucleic acid construct comprising as operably linked components in the 5′ to 3′ direction of transcription:
a transcriptional initiation region; and a polynucleotide sequence encoding an enzyme active in the formation of triacylglycerol from diacylglycerol and fatty acyl substrates.
- 23. The nucleic acid construct according to claim 22, wherein said enzyme is diacylglycerol acyltransferase.
- 24. A host cell comprising a DNA construct according to claim 22.
- 25. The host cell according to claim 24, wherein said host cell is selected from the group consisting of bacterial, insect, fungal, mammalian, and plant.
- 26. A plant comprising a cell of claim 25.
- 27. A method for producing a recombinant host cell, comprising:
transforming or transfecting a cell with a nucleic acid construct comprising a transcriptional initiation region and a polynucleotide sequence encoding an enzyme active in the formation of triacylglycerol from diacylglycerol and fatty acyl substrates, such that said host cell, under appropriate culture conditions, produces an acyltransferase protein.
- 28. The method according to claim 2723 wherein said host cell is a plant cell.
- 29. A method for producing a recombinant host cell, comprising:
transforming or transfecting a cell with a nucleic acid construct comprising a transcriptional initiation region and a polynucleotide sequence selected from the group consisting of a polynucleotide according to claim 12 and a polynucleotide according to claim 13, such that said host cell, under appropriate culture conditions, produces an acyltransferase protein.
- 30. The method according to claim 29, wherein said polynucleotide sequence comprises the nucleotide sequence set forth in SEQ ID NO: 37.
- 31. The method according to claim 29, wherein said host cell is a plant cell.
- 32. A method of modifying the triacylglycerol composition in a plant cell, said method comprising:
transforming a plant cell with a nucleic acid construct comprising a transcriptional initiation region and a polynucleotide sequence encoding an enzyme active in the formation of triacylglycerol from diacylglycerol and fatty acyl substrates, or fragment or variant thereof
- 33. The method according to claim 32, wherein said polynucleotide sequence is in an antisense orientation.
- 34. The method according to claim 32, wherein said polynucleotide sequence is in a sense orientation.
- 35. The method according to claim 32, wherein the activity of the endogenous diacylglycerol acyltransferase protein is suppressed.
- 36. The method according to claim 32, wherein the activity of the endogenous diacylglycerol acyltransferase protein is enhanced.
- 37. A method of modifying the lipid composition in a plant cell, said method comprising:
transforming a plant cell with a nucleic acid construct comprising a transcriptional initiation region and a polynucleotide sequence selected from the group consisting of a polynucleotide according to claim 12 and a polynucleotide according to claim 13, such that said plant cell, under appropriate culture conditions, produces an acyltransferase protein
- 38. The method according to claim 37 wherein said polynucleotide sequence is in an antisense orientation, whereby transcribed mRNA from said sequence is complementary to the equivalent mRNA transcribed from the endogenous gene and the activity of said diacylglycerol acyltransferase protein in said plant cell is suppressed.
- 39. The method according to claim 37, wherein said polynucleotide sequence is in a sense orientation.
- 40. An antibody immunospecific for the polypeptide of claim 10.
- 41. An antibody immunospecific for the polypeptide of claim 11.
- 42. A method of treating or ameliorating a disease or condition associated with altered diacylglycerol acyl transferase (DAGAT) activity, said method comprising the step of:
administering a therapeutically effective amount of an antagonist of DAGAT activity to a subject in need thereof.
- 43. A method of treating or ameliorating a disease or condition associated with altered diacylglycerol acyl transferase (DAGAT) activity, said method comprising the step of:
administering a therapeutically effective amount of an agonist of DAGAT activity to a subject in need thereof.
- 44. A method of diagnosing a disease or susceptibility to a disease associated with DAGAT expression or activity in a patient, said method comprising the step of:
determining the presence or absence of a mutation in the nucleotide sequence encoding DAGAT in the genome of said patient.
- 45. A method of diagnosing a disease or susceptibility to a disease associated with DAGAT expression or activity in a patient, said method comprising the step of:
detecting the presence or amount of DAGAT or an indicator thereof in a sample obtained from said patient.
- 46. A method for identifying a compound which stimulate or inhibit DAGAT expression or activity comprising:
contacting a composition comprising a polypeptide selected from the group consisting of a polypeptide comprising the amino acid sequence set forth in SEQ ID NO: 38 and a polypeptide comprising the amino acid sequence set forth in SEQ ID NO: 45 with a candidate compound; and detecting an interaction between said polypeptide and said candidate compound.
- 47. An agonist of a polypeptide selected from the group consisting of a polypeptide comprising the amino acid sequence set forth in SEQ ID NO: 38 and a polypeptide comprising the amino acid sequence set forth in SEQ ID NO: 45.
- 48. An antagonist of a polypeptide selected from the group consisting of a polypeptide comprising the amino acid sequence set forth in SEQ ID NO: 38 and a polypeptide comprising the amino acid sequence set forth in SEQ ID NO: 45.
Parent Case Info
[0001] This application claims the benefit of U.S. Provisional Application Number 60/091,631 filed Jul. 2, 1998, and U.S. Provisional Application No. 60/130,829 filed Apr. 23, 1999.
Provisional Applications (2)
|
Number |
Date |
Country |
|
60091631 |
Jul 1998 |
US |
|
60130829 |
Apr 1999 |
US |
Continuations (1)
|
Number |
Date |
Country |
Parent |
09345461 |
Jun 1999 |
US |
Child |
10121857 |
Apr 2002 |
US |