Nucleic acids encoding anthelmintic agents and plants made therefrom
원문보기
IPC분류정보
국가/구분
United States(US) Patent
등록
국제특허분류(IPC7판)
C12N-015/09
C12N-015/29
C12N-015/82
A01H-005/00
출원번호
US-0772227
(2004-02-04)
등록번호
US-7365240
(2008-04-29)
발명자
/ 주소
Verbsky,Michelle L.
Baublite,Catherine
Williams,Deryck J.
Kloek,Andrew P.
Davila Aponte,Jennifer A.
Hresko,Michelle Coutu
McLaird,Merry B.
Zentella,Rodolfo
출원인 / 주소
Divergence, Inc.
대리인 / 주소
Fish & Richardson P.C.
인용정보
피인용 횟수 :
0인용 특허 :
37
초록▼
The present invention provides DNA constructs, transgenic plants containing such constructs, and methods of making the plants. The DNA constructs encode a polypeptide that when expressed results in the production of fatty acid compounds having anthelmintic activity. Transgenic plants expressing suc
The present invention provides DNA constructs, transgenic plants containing such constructs, and methods of making the plants. The DNA constructs encode a polypeptide that when expressed results in the production of fatty acid compounds having anthelmintic activity. Transgenic plants expressing such a polypeptide can exhibit enhanced resistance to plant parasitic nematodes, particularly when expressed in vegetative tissues.
대표청구항▼
What is claimed is: 1. A transgenic plant containing at least one DNA construct, said construct comprising: a) a nucleic acid encoding a polypeptide effective for catalysing the conversion of a substrate to a C16, C18, or C20 monounsaturated fatty acid products wherein said polypeptide has at least
What is claimed is: 1. A transgenic plant containing at least one DNA construct, said construct comprising: a) a nucleic acid encoding a polypeptide effective for catalysing the conversion of a substrate to a C16, C18, or C20 monounsaturated fatty acid products wherein said polypeptide has at least 95% sequence identity to the amino acid sequence shown in SEQ ID NO:41, wherein said fatty acid product has the following structure: wherein X is hydrogen, CoA, glycerol, a monoglyceride, a diglyceride, ACP, methyl, Na+, phosphatidyicholine, or phosphatidylethanolamine, and wherein R3 is C2, C4, or C6 alkyl; and (b) a regulatory element operably linked to said nucleic acid encoding said polypeptide, wherein said regulatory element confers expression in a vegetative tissue of said plant; wherein the transgenic plant has anthelmintic activity. 2. The plant according to claim 1, wherein the double bond between the 9th and 10th carbons is cis. 3. The plant according to claim 1, wherein the double bond between the 9th and 10th carbons is trans. 4. The plant according to claim 1, wherein said regulatory element is a 5'-regulatory element. 5. The plant of claim 4, wherein said 5'-regulatory element confers expression in root tissue. 6. The plant of claim 5, said plant having a significantly increased amount of a epoxy-fatty acid in roots of said plant relative to a corresponding plant that lacks said DNA construct. 7. The plant of claim 6, wherein said epoxy-fatty acid is vernolic acid. 8. The plant of claim 7, wherein said vernolic acid constitutes from about 0.1% to about 35% of the total fatty acid content of said roots. 9. The plant of claim 4, wherein said 5'-regulatory element is selected from the group consisting of a potato ribosomal protein S27a Ubi3 promoter, a RB7 promoter, an alfalfa histone H3.2 promoter, an IRT2 promoter, an Arabidopsis FAD2 5'-UTR, an Arabidopsis FAD3 5'-UTR, a Ubi3 5'-UTR, an alfalfa histone H3.2 5'-UTR, and a CaMV35S 5'-UTR. 10. The plant of claim 1, wherein said regulatory element comprises a first 5'-regulatory element operably linked to a second 5'-regulatory element, wherein said first 5'-regulatory element is an Ubi3 promoter and said second 5'-regulatory element is selected from the group consisting of an Arabidopsis FAD2 5'-UTR, an Arabidopsis FAD3 5'-UTR, a potato ribosomal protein S27a 5'-UTR, a Ubi3 5'-UTR, and a CaMV35S 5'-UTR. 11. The plant of claim 4, wherein said DNA construct further comprises a 3'-regulatory element. 12. The plant of claim 11, wherein said 3'-regulatory element comprises a Ubi3 terminator or an E9 pea terminator. 13. The plant of claim 11, wherein said 5'-regulatory element is selected from the group consisting of an Arabidopsis FAD2 5'-UTR and an Arabidopsis FAD3 5'-UTR and said 3'-regulatory element is selected from the group consisting of an Arabidopsis FAD2 3'-UTR and an Arabidopsis FAD3 3'-UTR. 14. The plant of claim 13, wherein said 5'-regulatory element comprises SEQ ID NOS: 43 or 44 and said 3'-regulatory element comprises SEQ ID NO: 45. 15. The plant of claim 1, wherein said at least one DNA construct further comprises at least one regulatory element that confers expression in vegetative tissues of a plant operably linked to a nucleic acid that encodes a PDAT or DAGAT polypeptide. 16. The plant of claim 1, said plant further comprising a second DNA construct, said second DNA construct comprising at least one regulatory element that confers expression in vegetative tissues of a plant operably linked to a nucleic acid that encodes a PDAT or DAGAT polypeptide. 17. The plant of claim 1, wherein R3 is C2 alkyl or C4 alkyl. 18. The plant of claim 1, wherein said nucleic acid encoding said polypeptide has the nucleotide sequence shown in SEQ ID NO: 32. 19. The plant of claim 1, where said plant is selected from the group consisting of tobacco, tomato, soybean, corn, cotton, rice, wheat, banana, carrot, potato, strawberry and turf grass. 20. A method of making a transgenic plant having anthelmintic activity, said method comprising introducing a construct into a plant, wherein said construct comprises: a) a nucleic acid encoding a polypeptide effective for catalysing the conversion of a substrate to a C16, C18, or C20 monounsaturated fatty acid product, wherein said polypeptide has the amino acid sequence shown in SEQ ID NO:41, wherein said fatty acid product has the following structure: wherein X is hydrogen, CoA, glycerol, a monoglyceride, a diglyceride, ACP, methyl, Na+, phosphatidyicholine, or phosphatidylethanolamine, and wherein R3 is C2, C4, or C6 alkyl; and (b) a regulatory element operably linked to said nucleic acid encoding said polypeptide, wherein said regulatory element confers expression in a vegetative tissue of said plant. 21. The method of claim 20, wherein said regulatory element of said construct is a 5'-regulatory element. 22. The method of claim 21, wherein said 5'-regulatory element comprises a potato ribosomal protein S27a Ubi3 promoter, a RB7 promoter, an alfalfa histone H3.2 promoter, an IRT2 promoter, an Arabidopsis FAD2 5'-UTR, an Arabidopsis FAD3 5'-UTR, a Ubi3 5'-UTR, an alfalfa histone H3.2 5'-UTR, and a CaMV35S 5'-UTR. 23. The method of claim 21, wherein said regulatory element comprises a first 5'-regulatory element operably linked to a second 5'-regulatory element, wherein said first 5'-regulatory element is an Ubi3 promoter and said second 5'-regulatory element is selected from the group consisting of an Arabidopsis FAD2 5'-UTR, an Arabidopsis FAD3 5'-UTR, a potato ribosomal protein S27a 5'-UTR, a Ubi3 5'-UTR, and a CaMV35S 5'-UTR. 24. The method of claim 21, wherein said DNA construct further comprises a 3'-regulatory element. 25. The method of claim 24, wherein said 5'-regulatory element comprises SEQ ID NO: 43 or SEQ ID NO: 44 and said 3'-UTR comprises SEQ ID NO: 45. 26. An isolated nucleic acid comprising the nucleotide sequence set forth in SEQ ID NO:32. 27. A recombinant nucleic acid construct comprising at least one regulatory element that confers expression in a vegetative tissue of a plant, said regulatory element operably linked to a nucleic acid having the nucleotide sequence shown in SEQ ID NO:32. 28. The nucleic acid construct of claim 27, wherein said at least one regulatory element comprises a 5'-regulatory element having the nucleotide sequence set forth in SEQ ID NO: 43 or SEQ ID NO: 44. 29. The nucleic acid construct of claim 28, wherein said construct further comprises a 3'-regulatory element having the nucleotide sequence set forth in SEQ ID NO: 45. 30. A transgenic plant harboring a DNA construct comprising a nucleic acid encoding a fatty acid epoxygenase polypeptide operably linked to a regulatory element conferring expression of said polypeptide in a vegetative tissue of said plant, wherein said polypeptide has the amino acid sequence shown in SEQ ID NO:41. 31. The plant of claim 30, said plant having a significantly increased amount of a epoxy-fatty acid in roots of said plant relative to a corresponding plant that lacks said DNA construct. 32. The plant of claim 31, wherein said epoxy-fatty acid is vemolic acid. 33. The plant of claim 32, wherein said vernolic acid constitutes from about 0.1% to about 35% of the total fatty acid content of said roots.
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