VCP-Based Vectors for Algal Cell Transformation

• US 20090317904A1
• Filed: 06/08/2009
• Published: 12/24/2009
• Est. Priority Date: 06/06/2008
• Status: Active Grant

• Current Assignee: Aurora Algae, Incorporated (Reliance Industries Limited)
• Original Assignee: Aurora Algae, Incorporated (Reliance Industries Limited)
• Inventors: Kilian, Oliver, Vick, Bertrand
• Granted Patent: US 8,318,482 B2
• Time in Patent Office: Days
• US Class Current: 435/320.100
• CPC Class Codes: C12N 1/12 Unicellular algae; Culture media therefor as new plants A01H13/00
• CPC Class Codes: C12N 15/79 Vectors or expression systems specially adapted for eukaryotic hosts
• CPC Class Codes: C12N 15/8206 by physical or chemical, i.e. non-biological, means, e.g. electroporation, PEG mediated
• CPC Class Codes: C12N 15/8207 by mechanical means, e.g. microinjection, particle bombardment, silicon whiskers
• CPC Class Codes: C12N 15/8237 Externally regulated expression systems
• CPC Class Codes: C12N 15/8247 involving modified lipid metabolism, e.g. seed oil composition
• CPC Class Codes: C12N 15/90 Stable introduction of foreign DNA into chromosome

Abstract

Provided herein are exemplary vectors for transforming algal cells. In exemplary embodiments, the vector comprises a Violaxanthin-chlorophyll a binding protein (Vcp) promoter driving expression of an antibiotic resistance gene in an algal cell. Embodiments of the invention may be used to introduce a gene (or genes) into the alga Nannochloropsis, such that the gene(s) are expressed and functional. This unprecedented ability to transform Nannochloropsis with high efficiency makes possible new developments in phycology, aquaculture and biofuels applications.

Claims

1. An expression vector for algal cell transformation comprising a nucleotide sequence shown in SEQ. ID. NO. 1.

2. The expression vector of claim 1, wherein the nucleotide sequence is at least ninety percent (90%) similar to the nucleotide sequence shown in SEQ. ID. NO. 1.

3. The expression vector of claim 1, wherein a promoter in the expression vector modulates the expression of a gene governed by the promoter in response to light intensity.

4. The expression vector of claim 3, wherein the promoter in the expression vector increases the expression of the gene governed by the promoter in response to a decrease in light intensity.

5. The expression vector of claim 3, wherein the promoter is a Vcp promoter.

6. The expression vector of claim 1, wherein the algal cell is of algal genus Nannochloropsis.

7. The expression vector of claim 1, wherein the expression vector is at least partially expressed as part of a transformed algal cell.

8. An expression vector for algal cell transformation comprising a nucleotide sequence shown in SEQ. ID. NO. 2.

9. The expression vector of claim 8, wherein the nucleotide sequence is at least ninety percent (90%) similar to the nucleotide sequence shown in SEQ. ID. NO. 2.

10. The expression vector of claim 8, wherein a promoter in the expression vector modulates the expression of a gene governed by the promoter in response to light intensity.

11. The expression vector of claim 10, wherein the promoter in the expression vector increases the expression of the gene governed by the promoter in response to a decrease in light intensity.

12. The expression vector of claim 10, wherein the promoter is a Vcp promoter.

13. The expression vector of claim 8, wherein the algal cell is of algal genus Nannochloropsis.

14. The expression vector of claim 8, wherein the expression vector is at least partially expressed as part of a transformed algal cell.

15. An expression vector for algal cell transformation comprising a nucleotide sequence shown in SEQ. ID. NO. 3.

16. The expression vector of claim 15, wherein the nucleotide sequence is at least ninety percent (90%) similar to the nucleotide sequence shown in SEQ. ID. NO. 3.

17. The expression vector of claim 15, wherein a promoter in the expression vector modulates the expression of a gene governed by the promoter in response to light intensity.

18. The expression vector of claim 17, wherein the promoter in the expression vector increases the expression of the gene governed by the promoter in response to a decrease in light intensity.

19. The expression vector of claim 17, wherein the promoter is a Vcp promoter.

20. The expression vector of claim 15, wherein the algal cell is of algal genus Nannochloropsis.

21. The expression vector of claim 15, wherein the expression vector is at least partially expressed as part of a transformed algal cell.