Claims
- 1. An isolated nucleic acid molecule which encodes a polypeptide having an activity of catalyzing the formation of a double bond, wherein said nucleic acid molecule comprises:
a. a nucleotide sequence of LuFad3 from the genus Linum; b. a nucleotide sequence which is at least about 60% identical to the nucleotide sequence of SEQ ID NO:7, or a complement thereof; c. a nucleotide sequence comprising a fragment of the nucleotide sequence of SEQ ID NO:7; d. a nucleotide sequence which encodes a polypeptide comprising an amino acid sequence that is at least about 70% homologous to the amino acid sequence of SEQ ID NO:8; e. a nucleotide sequence which encodes a fragment of a polypeptide comprising the amino acid sequence of SEQ ID NO:8, wherein the fragment comprises at least 15 contiguous amino acids of SEQ ID NO:8; or f. a nucleotide sequence which encodes a naturally occurring allelic variant of a polypeptide comprising the amino acid sequence of SEQ ID NO:8, wherein the nucleic acid molecule hybridizes to a nucleic acid molecule comprising SEQ ID NO:7, or a complement thereof under stringent conditions.
- 2. The isolated nucleic acid molecule of claim 1, wherein the nucleic acid molecule encodes a polypeptide having an activity of catalyzing the formation of a double bond at position 15 from the carboxyl end of a fatty acyl chain.
- 3. An isolated nucleic acid molecule, wherein said nucleic acid molecule comprises:
a. a nucleotide sequence of Conlinin 1 from the genus Linum; b. a nucleotide sequence which is at least about 60% identical to the nucleotide sequence of SEQ ID NO:1, or a complement thereof; c. a nucleotide sequence comprising a fragment of the nucleotide sequence of SEQ ID NO:1; d. a nucleotide sequence which encodes a polypeptide comprising an amino acid sequence that is at least about 60% homologous to the amino acid sequence of SEQ ID NO:2; e. a nucleotide sequence which encodes a fragment of a polypeptide comprising the amino acid sequence of SEQ ID NO:2, wherein the fragment comprises at least 15 contiguous amino acids of SEQ ID NO:2; or f. a nucleotide sequence which encodes a naturally occurring allelic variant of a polypeptide comprising the amino acid sequence of SEQ ID NO:2, wherein the nucleic acid molecule hybridizes to a nucleic acid molecule comprising SEQ ID NO:1, or a complement thereof under stringent conditions.
- 4. An isolated nucleic acid molecule, wherein said nucleic acid molecule comprises:
a. a nucleotide sequence of Conlinin 2 from the genus Linum; b. a nucleotide sequence which is at least about 60% identical to the nucleotide sequence of SEQ ID NO:3, or a complement thereof; c. a nucleotide sequence comprising a fragment of the nucleotide sequence of SEQ ID NO:3; d. a nucleotide sequence which encodes a polypeptide comprising an amino acid sequence that is at least about 55% homologous to the amino acid sequence of SEQ ID NO:4; e. a nucleotide sequence which encodes a fragment of a polypeptide comprising the amino acid sequence of SEQ ID NO:4, wherein the fragment comprises at least 15 contiguous amino acids of SEQ ID NO:4; or f. a nucleotide sequence which encodes a naturally occurring allelic variant of a polypeptide comprising the amino acid sequence of SEQ ID NO:4, wherein the nucleic acid molecule hybridizes to a nucleic acid molecule comprising SEQ ID NO:3, or a complement thereof under stringent conditions.
- 5. A vector comprising the nucleic acid molecule of any one of claims 1, 3, or 4.
- 6. A host cell transformed with the vector of claim 5.
- 7. The vector of claim 5, which is an expression vector.
- 8. A host cell transformed with the vector of claim 7.
- 9. A method of producing a polypeptide comprising culturing the host cell of claim 8 in an appropriate culture medium to, thereby, produce the polypeptide.
- 10. The host cell of claim 6, wherein the cell is a plant cell.
- 11. A plant transformed with the nucleic acid molecule of any one of claims 1, 3, or 4.
- 12. A method of producing a cell capable of generating α-linoleic acid, said method comprising introducing into said cell the nucleic acid molecule of claim 1, wherein the nucleic acid molecule encodes a desaturase having an activity of catalyzing the formation of a double bond at position 15 from the carboxyl end of a fatty acyl chain.
- 13. A promoter comprising a nucleotide sequence isolated from Linum which is capable of directing gene expression in developing flax seeds.
- 14. An isolated Linum Conlinin 1 promoter comprising the nucleotide sequence of SEQ ID NO: 5, or a portion thereof.
- 15. An isolated Linum Conlinin 2 promoter comprising the nucleotide sequence of SEQ ID NO: 6, or a portion thereof.
- 16. A vector comprising the Linum Conlinin promoter of claim 14 or claim 15 operably linked to a heterologous gene of interest.
- 17. An isolated Linum LuFad 3 promoter comprising the nucleotide sequence of SEQ ID NO: 9, or a portion thereof
- 18. The promoter of claim 17, wherein the Linum type is cultivar CDC Normandy.
- 19. An isolated Linum LuFad3 promoter comprising the nucleotide sequence of SEQ ID NO: 10, or a portion thereof.
- 20. The promoter of claim 19, wherein the Linum type is cultivar CDC Solin.
- 21. A vector comprising a Linum LuFad3 promoter of claims 17 or 19, operably linked to a heterologous gene of interest.
- 22. An isolated nucleic acid sequence capable of directing seed-specific expression in a plant comprising
a. a nucleic acid comprising the nucleotides of SEQ ID NO: 5 or SEQ ID NO: 6; b. a nucleic acid sequence that is complimentary to the nucleotide sequence of SEQ ID NO: 5 or SEQ ID NO: 6; or c. a nucleic acid sequence that is at least about 60% homologous to the nucleotide sequence of SEQ ID NO: 5 or SEQ ID NO: 6.
- 23. An isolated nucleic acid sequence capable of directing seed-specific expression in a plant comprising
a. a nucleic acid comprising the nucleotides of SEQ ID NO: 9 or SEQ ID NO: 10; b. a nucleic acid sequence that is complimentary to the nucleotide sequence of SEQ ID NO: 9 or SEQ ID NO: 10; or c. a nucleic acid sequence that is at least about 60% homologous to the nucleotide sequence of SEQ ID NO: 9 or SEQ ID NO: 10.
- 24. A method for the expression of a nucleic acid sequence of interest in flax seeds comprising;
a. preparing a nucleic acid construct comprising a seed-specific promoter from any of claims 14, 15, 17, or 19 operably linked to a gene of interest, wherein the gene of interest is non-native to the seed-specific promoter; b. introducing said construct into a flax plant cell; and c. growing said cell into a mature plant capable of setting seed wherein the gene of interest is expressed in the seed under the control of the seed-specific promoter.
- 25. A transgenic flax plant prepared by the method of claim 24.
- 26. A transgenic seed having a transgene integrated into the genome of the seed, wherein the transgene comprises a seed-specific promoter from any of claims 14, 15, 17, or 9 operatively linked to a gene of interest, wherein the promoter-linked gene of interest confers a detectable and functional phenotype on the seed when expressed.
- 27. A promoter comprising a nucleotide sequence isolated from Linum which is capable of directing gene expression in developing flax seeds operatively linked to a gene related to fatty acid biosythesis or lipid biosynthesis.
- 28. The promoter of claim 27 which is selected from the group consisting of SBQ ID NO:5, SEQ ID NO:6, SEQ ID NO:9, or SEQ ID NO:10.
- 29. The promoter of claim 27, wherein the gene of interest is selected from the group consisting of a conjugase, Δ4 desaturase, Δ5 desaturase, or Δ6 desaturase.
- 30. A promoter capable of directing gene expression in developing flax seeds operatively linked to a gene related to fatty acid biosythesis or lipid biosynthesis, wherein the promoter comprises at least 701 nucleotides of SEQ ID NO:5, 751 nucleotides of SEQ ID NO:5, 801 nucleotides of SEQ ID NO:5, 851 nucleotides of SEQ ID NO:5, 901 nucleotides of SEQ ID NO:5, 951 nucleotides of SEQ ID NO:5, 1,001 nucleotides of SEQ ID NO:5, or 1,118 nucleotides of SEQ ID NO:5.
- 31. A promoter capable of directing gene expression in developing flax seeds operatively linked to a gene related to fatty acid biosythesis or lipid biosynthesis, wherein the promoter comprises at least 597 nucleotides of SEQ ID NO:6, 647 nucleotides of SEQ ID NO:6, 697 nucleotides of SEQ ID NO:6, 746 nucleotides of SEQ ID NO:6, 797 nucleotides of SEQ ID NO:6, 846 nucleotides of SEQ ID NO:6m 897 nucleotides of SEQ ID NO:6, 947 nucleotides of SEQ ID NO:6, 997 nucleotides of SEQ ID NO:6, or 1,014 nucleotides of SEQ ID NO:6.
- 32. The promoter, or portion thereof, from the Conlinin 1 gene that is not 417 nucleotides in length.
- 33. The promoter, or portion thereof, from the Conlinin 2 gene that is not 417 nucleotides in length.
- 34. The host cell of claim 8, wherein the cell is a plant cell.
RELATED APPLICATIONS
[0001] This application claims priority to U.S. Provisional Application No.: 60/295,823 entitled “Flax (Linum usitatissimum L.) Speed-Specific Promoters” filed Jun. 6, 2001, the entire contents of which are hereby incorporated by reference. The entire contents of Appendix A including the entire contents of all references cited therein also are expressly incorporated by reference and are intended to be part of the present application.
Provisional Applications (1)
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Number |
Date |
Country |
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60295823 |
Jun 2001 |
US |