Desaturase genes and their use

• US 6,897,050 B1
• Filed: 11/24/1998
• Issued: 05/24/2005
• Est. Priority Date: 11/24/1997
• Status: Expired due to Fees

• Current Assignee: University of Bristol
• Original Assignee: University of Bristol
• Inventors: Napier, Johnathan A.
• Primary Examiner(s): Prouty, Rebecca E.
• Assistant Examiner(s): Walicka, Malgorzata A.
• Application Number:
  US09/555,093
• Time in Patent Office: 2,373 Days
• Field of Search: 435/189, 435/226, 530/350
• US Class Current: 435/189
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61P 1/16 for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
• CPC Class Codes: A61P 17/10 Anti-acne agents
• CPC Class Codes: A61P 29/00 Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 31/12 Antivirals
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 9/10 for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
• CPC Class Codes: A61P 9/12 Antihypertensives
• CPC Class Codes: C12N 15/8209 Selection, visualisation of transformants, reporter constructs, e.g. antibiotic resistance markers
• CPC Class Codes: C12N 15/8247 involving modified lipid metabolism, e.g. seed oil composition
• CPC Class Codes: C12N 15/8271 for stress resistance, e.g. heavy metal resistance
• CPC Class Codes: C12N 9/0083 Miscellaneous (1.14.99)
• 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 7/6472 containing polyunsaturated fatty acid [PUFA] residues, i.e. having two or more double bonds in their backbone

Abstract

cDNA encoding C. elegans Δ⁶ desaturase has been cloned and sequenced, and the Δ⁶ desaturase amino acid sequence has been determined: The C. elegans Δ⁶ desaturase has a surprisingly low level of sequence identity with the known borage Δ⁶ desaturase. The C. elegans Δ⁶ desaturase has been expressed in yeast. It and other desaturases can be cloned in host organisms (e.g. plants) and can be used to provide useful metabolites.

Claims

1. An isolated and purified polypeptide having Δ

⁶ desaturase activity which comprises an amino acid sequence that is at least 95% identical to SEQ ID NO;

2.

2. A polypeptide according to claim 1, which has a cytochrome b₅ domain.

3. A polypeptide according to claim 1, which comprises a histidine box.

4. A polypeptide according to claim 1, which occurs naturally in C. elegans.

5. A polypeptide according to claim 1, which is covalently linked to a moiety capable of being isolated by affinity chromatography.

9. The polypeptide of claim 1 which is covalently linked to a moiety capable of being isolated by affinity chromatography.

11. The polypeptide of claim 1 which has at least 99% amino acid sequence identity with the amino acid sequence of SEQ ID NO:

2.

6. An isolated and purified Δ

⁶ desaturase polypeptide comprising the amino acid sequence as shown in SEQ ID NO;

2 with no more than five conservative amino acid substitutions.

7. The polypeptide of claim 6 which has one conservative amino acid substitution.

8. The polypeptide of claim 6 which comprises the amino acid sequence as shown in SEQ ID NO:

2.

10. The polypeptide of claim 8 further comprising a signal sequence.

12. An isolated and purified polypeptide having Δ

⁶ desaturase activity, which comprises a fragment of SEQ ID NO;

2, wherein said fragment has Δ

⁶ desaturase activity.