Nova Patents
CA2625855C

Delta-9 elongases and their use in making polyunsaturated fatty acids

Abstract

Isolated nucleic acid fragments and recombinant constructs comprising such fragments encoding delta-9 elongases along with a method of making long-chain polyunsaturated fatty acids (PUFAs) using these delta-9 elongases in plants.

CA2625855C, drawing sheet 1
Sheet 1 of 16

Term

0.1 yearsleft in the term

Expires 16 November 2026.

  1. Priority
  2. Filed
  3. Granted
  4. Today
  5. Expires

20 claims: 1 independent, 19 dependent

  1. 1
    CA 02625855 2015-01-23 CLAIMS What is claimed is:1. A method for producing at least one polyunsaturated fatty acid in a soybean cell comprising: (a) transforming a soybean cell with a first recombinant construct comprising an isolated polynucleotide comprising: (i) a nucleotide sequence encoding a polypeptide having delta9 elongase activity, wherein the polypeptide has at least 70% amino acid identity, based on the Clustal V method of alignment, when compared to the amino acid sequence as set forth in SEQ ID NO:5;(ii) a nucleotide sequence encoding a polypeptide having delta9 elongase activity, wherein the nucleotide sequence has at least 70% sequence identity, based on the BLASTN method of alignment, when compared to the nucleotide sequence as set forth in SEQ ID NO:4 or SEQ ID NO:90;(iii) a nucleotide sequence encoding a polypeptide having delta9 elongase activity, wherein the nucleotide sequence hybridizes with a complement of the nucleotide sequence as set forth in SEQ ID NO:4 or SEQ ID NO:90 under the following stringent hybridization conditions: 0.1X SSC, 0.1% SDS, 65 °C and washed with 2X SSC, 0.1% SDS followed byO.IXSSC, 0.1% SDS;or (iv) a complement of the nucleotide sequence of (i), (ii) or (iii), wherein the complement and the nucleotide sequence consist of the same number of nucleotides and are 100% complementary;operably linked to at least one regulatory sequence and at least one additional recombinant construct comprising an isolated polynucleotide, operably linked to at least one regulatory sequence, encoding a polypeptide that is a delta-4 desaturase, a delta-5 desaturase, a delta-6 desaturase, a delta-12 desaturase, a 130 CA 02625855 2015-01-23 delta-15 desaturase, a delta-17 desaturase, a delta-8 desaturase, a C14/16 elongase, a Ci6/ia elongase, a C18/20 elongase or a C20/22 elongase;(b) regenerating a soybean plant from the transformed soybean cell of step (a);and (c) selecting those seeds obtained from the plant of step (b) having an altered level of polyunsaturated fatty acids when compared to the level in seeds obtained from a nontransformed soybean plant. An oilseed plant cell comprising: (a) a first recombinant construct comprising an isolated polynucleotide comprising: (i) a nucleotide sequence encoding a polypeptide having delta9 elongase activity, wherein the polypeptide has at least 70% amino acid identity, based on the Clustal V method of alignment, when compared to the amino acid sequence as set forth in SEQ ID NO:5;(ii) a nucleotide sequence encoding a polypeptide having delta9 elongase activity, wherein the nucleotide sequence has at least 70% sequence identity, based on the BLASTN method of alignment, when compared to the nucleotide sequence as set forth in SEQ ID NO:4 or SEQ ID NO:90;(iii) a nucleotide sequence encoding a polypeptide having delta9 elongase activity, wherein the nucleotide sequence hybridizes with a complement of the nucleotide sequence as set forth in SEQ ID NO:4 or SEQ ID NO:90 under the following stringent hybridization conditions: 0.1X SSC, 0.1% SDS, 65 °C and washed with 2X SSC, 0.1% SDS followed byO.IXSSC, 0.1% SDS;or (iv) a complement of the nucleotide sequence of (i), (ii) or (iii), wherein the complement and the nucleotide sequence consist of the same number of nucleotides and are 100% complementary;131 CA 02625855 2015-01-23 operably linked to at least one regulatory sequence;and (b) at least one additional recombinant construct comprising an isolated polynucleotide, operably linked to at least one regulatory sequence, encoding a polypeptide that is a delta-4 desaturase, a 5 delta-5 desaturase, a delta-6 desaturase, a delta-12 desaturase, a delta-15 desaturase, a delta-17 desaturase, a delta-8 desaturase, a C14/16 elongase, a C16/18 elongase, a C18/20 elongase or a C20/22 elongase.
  2. 2
    3. The oilseed plant cell of claim 2, wherein the oilseed plant cell is from 10 soybean, Brassica species, sunflower, maize, cotton, flax or safflower.
  3. 6
    7. The method of claim 6, wherein the polypeptide of (a)(i) has at least 90% amino acid identity, based on the Clustal V method of alignment, when compared to the amino acid sequence as set forth in SEQ ID NO:5. 25
  4. 7
    8. The method of claim 7, wherein the polypeptide of (a)(i) has at least 95% amino acid identity, based on the Clustal V method of alignment, when compared to the amino acid sequence as set forth in SEQ ID NO:5.
  5. 8
    9. The method of claim 8, wherein the polypeptide of (a)(i) comprises the amino acid sequence set forth in SEQ ID NO:5. 132 CA 02625855 2015-01-23
  6. 10
    11. The method of claim 10, wherein the nucleotide sequence of (a)(ii) has at least 90% sequence identity, based on the BLASTN method of alignment, when compared to the nucleotide sequence as set forth in SEQ ID NO:4 or SEQ ID NO:90.
  7. 11
    12. The method of claim 11, wherein the nucleotide sequence of (a)(ii) has at 10 least 95% sequence identity, based on the BLASTN method of alignment, when compared to the nucleotide sequence as set forth in SEQ ID NO:4 or SEQ ID NO:90.
  8. 12
    13. The method of claim 12, wherein the nucleotide sequence of (a)(ii) comprises the nucleotide sequence set forth in SEQ ID NO:4 or SEQ ID 15 NO:90.
  9. 14
    15. The oilseed plant cell of claim 14, wherein the polypeptide of (a)(i) has at least 90% amino acid identity, based on the Clustal V method of alignment, when compared to the amino acid sequence as set forth in SEQ ID NO:5.
  10. 15
    16. The oilseed plant cell of claim 15, wherein the polypeptide of (a)(i) has at 25 least 95% amino acid identity, based on the Clustal V method of alignment, when compared to the amino acid sequence as set forth in SEQ ID NO:5.
  11. 16
    17. The oilseed plant cell of claim 16, wherein the polypeptide of (a)(i) comprises the amino acid sequence set forth in SEQ ID NO:5. 133 CA 02625855 2015-01-23
  12. 18
    19. The oilseed plant cell of claim 18, wherein the nucleotide sequence of (a)(ii) has at least 90% sequence identity, based on the BLASTN method of alignment, when compared to the nucleotide sequence as set forth in SEQ ID NO:4 or SEQ ID NO:90.
  13. 19
    20. The oilseed plant cell of claim 19, wherein the nucleotide sequence of 10 (a)(ii) has at least 95% sequence identity, based on the BLASTN method of alignment, when compared to the nucleotide sequence as set forth in SEQ ID NO:4 or SEQ ID NO:90.
  14. 20
    21. The oilseed plant cell of claim 20, wherein the nucleotide sequence of (a)(ii) comprises the nucleotide sequence set forth in SEQ ID NO:4 or 15 SEQ ID NO:90. 134
Independent claims14