Transgenic cells

• US 20070061921A1
• Filed: 07/13/2004
• Published: 03/15/2007
• Est. Priority Date: 07/16/2003
• Status: Abandoned Application

• Current Assignee: University of York
• Original Assignee: University of York
• Inventors: Graham, Ian, Tonon, Thierry
• Application Number:
  US10/564,560
• Publication Number:
  US 20070061921A1
• Time in Patent Office: Days
• US Class Current: 800/281
• CPC Class Codes: A61P 3/02 Nutrients, e.g. vitamins, minerals
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• CPC Class Codes: C12N 15/8247 involving modified lipid metabolism, e.g. seed oil composition
• CPC Class Codes: C12N 9/1029 transferring groups other than amino-acyl groups (2.3.1)
• CPC Class Codes: C12P 7/6427 Polyunsaturated fatty acids [PUFA], i.e. having two or more double bonds in their backbone
• CPC Class Codes: C12P 7/6434 Docosahexenoic acids [DHA]
• CPC Class Codes: C12P 7/6472 containing polyunsaturated fatty acid [PUFA] residues, i.e. having two or more double bonds in their backbone

Abstract

We describe transgenic cells comprising nucleic acid molecules that comprise nucleic acid sequences which encode enzymes involved in the biosynthesis of n-3 fatty acids.

Claims

1. A transgenic cell comprising a nucleic acid molecule selected from the group consisting of:

i) a DNA molecule consisting of a DNA sequence as represented in SEQ ID NO;

1, 2, 3, or 4;

ii) a DNA molecule which hybridises to the sequences identified in (i) above and which encode a polypeptide which has fatty acid elongase activity; and

iii) DNA molecules consisting of DNA sequences that are degenerate as a result of the genetic code to the DNA sequence defined in (i) and (ii).

2. The cell according to claim 1 wherein said nucleic acid molecule anneals under stringent hybridisation conditions to the sequences described in (i), (ii) and (iii) above.

3. The cell according to claim 1 wherein said nucleic acid molecules are isolated from an algal species.

4. The cell according to claim 3 wherein said algal species is Amphidinium carterae, Amphiphora hyalina, Amphiphora sp., Chaetoceros gracilis, Coscinodiscus sp., Crypthecodinium cohnii, Cryptomonas sp., Cylindrotheca fusiformis, Haslea ostrearia, Isochrysis galbana, Nannochloropsis oculata, Navicula sp., Nitzschia closterium, Pavlova lutheri, Phaeodactylum tricornutum, Prorocentrum minimum, Rhizosolenia setigera, Skeletonema costatum, Skeletonema sp., Tetraselmis tetrathele, Thalassiosira nitzschioides, Thalassiosira heterophorma, Thalassiosira pseudonana, or Thalassiosira stellaris.

5. The cell of claim 1 wherein said polypeptide is a variant polypeptide and comprises the amino acid sequence shown in SEQ ID NO:

5, 6, or 7 which sequence has been modified by deletion, addition or substitution of at least one amino acid residue wherein said modification enhances the enzyme activity of said polypeptide.

6. The cell according to claim 5 wherein said modified polypeptide has enhanced fatty acid elongase activity

7. The cell according to claim 1 wherein said polypeptide comprises the amino acid sequence represented in SEQ ID NO:

5, 6, or 7.

8. The cell according to claim 7 wherein said polypeptide consists of the amino acid sequence represented in SEQ ID NO:

5, 6, or 7.

9. The cell according to any of claim 1 wherein said cell is transfected with a nucleic acid molecules selected from the group consisting of:

i) a DNA molecule consisting of the DNA sequence as represented in SEQ ID NO;

1, 2, 3, or 4;

ii) DNA molecules which hybridise to the sequences identified in (i) above and which encode a polypeptide which has fatty acid elongase activity; and

iii) DNA molecules comprising DNA sequences that are degenerate as a result of the genetic code to the DNA sequence defined in (i) and (ii);

combined with at least one of the nucleic acid molecules selected from the group consisting of;

iv) DNA molecules consisting of DNA sequences as represented in SEQ ID NO;

8, 10, 12, or 14;

v) DNA molecules which hybridise to the sequences identified in (i) above and which have desaturase, acyl-CoA synthetase or diacylglycerol acyltransferase activity;

vi) DNA molecules comprising DNA sequences that are degenerate as a result of the genetic code to the DNA sequence defined in (iv) and (v) above.

10. The cell according to claim 9 wherein said cell is a plant cell.

11. A plant comprising the cell of claim 9.

12. A seed comprising the cell of claim 9.

13. A foodstuff product comprising the cell of claim 9.

14. The foodstuff product of claim 13, wherein said foodstuff is wine;

beer;

bread;

baking products;

or vegetable extracts.

15. The food stuff according to claim 13 wherein said foodstuff is wine or beer.

16. A fermentation process comprising the cell of claim 9.

17. The fermentation process of claim 16 comprising:

providing a vessel containing the cell and constituents required for fermentation and fatty acid biosynthesis; and

providing conditions conducive to the fermentation of a liquid composition contained in said vessel.

18. An animal feed product comprising the cell of claim 9.

19. A method of modulating the level of n-3 fatty acid in a plant cell comprising;

providing a plant cell according to claim 10;

regenerating the plant cell into a plant; and

monitoring n-3 fatty acid production by said plant.

20. A method for the production and optionally the extraction of n-3 fatty acids comprising:

providing a cell according to claim 1;

providing conditions conducive to the growth of said cell; and

extracting n-3 fatty acids, or variants thereof, from said cell.

21. A method for the production and optionally the extraction of n-3 fatty acid comprising:

providing a plant cell according to claim 10;

regenerating said cell into a plant; and

extracting n-3 fatty acids, or variants thereof from said plant.

22. A reaction vessel comprising the cell of claim 1, fatty acid substrates and co-factors characterised in that said vessel is adapted for the conversion of said fatty acids substrates to n-3 fatty acids.