THERMOSTABLE DNA POLYMERASE FROM PALAEOCOCCUS HELGESONII

• US 20110104761A1
• Filed: 03/13/2009
• Published: 05/05/2011
• Est. Priority Date: 03/14/2008
• Status: Abandoned Application

• Current Assignee: Genesys Biotech Ltd.
• Original Assignee: Genesys Logic
• Inventors: Morant, Nicholas, Clark, Duncan
• Application Number:
  US12/922,401
• Publication Number:
  US 20110104761A1
• Time in Patent Office: Days
• US Class Current: 435/91.2
• CPC Class Codes: C12N 9/1252 DNA-directed DNA polymerase (2.7.7.7), i.e. DNA replicase

Abstract

There is provided a polypeptide having thermostable DNA polymerase activity and comprising or consisting of an amino acid sequence with at least 78% identity to Palaeococcus helgesonii DNA polymerase shown in SEQ ID NO: 1 or SEQ ID NO:39.

Claims

1. A polypeptide having thermostable DNA polymerase activity and comprising or consisting of an amino acid sequence with at least 79% identity to Palaeococcus helgesonii DNA polymerase shown in SEQ ID NO:

1.

2. The polypeptide according to claim 1 comprising or consisting of an amino acid sequence with at least 79% identity to Palaeococcus helgesonii DNA polymerase shown in SEQ ID NO:

39.

3. The polypeptide according to claim 1, which is suitable for carrying out a thermocycling amplification reaction, such as a polymerase chain reaction (PCR).

4. The polypeptide according to claim 1, in which the polypeptide has 3′

-5′

exonuclease proofreading activity.

5. The polypeptide according to claim 1, in which the polypeptide lacks 5′

-3′

exonuclease activity.

6. The polypeptide according to claim 1, which is an isolated thermostable DNA polymerase obtainable from Palaeococcus helgesonii and having a molecular weight of about 90,000 Daltons, or an enzymatically active fragment thereof.

7. A polypeptide according to claim 1 having thermostable DNA polymerase activity and comprising the amino acid sequence SEQ ID NO:

39.

9. A polypeptide according to claim 1, further comprising a Cren7 enhancer domain.

10. A composition comprising the polypeptide of claim 1.

11. The composition according to claim 10, which is enzymatically thermostable.

12. An isolated nucleic acid encoding the polypeptide of claim 1.

15. A vector comprising the nucleic acid of claim 12.

16. A host cell transformed with the nucleic acid of claim 12.

17. A kit comprising the polypeptide of claim 1, together with packaging materials therefor.

18. A method of amplifying a sequence of a target nucleic acid using a thermocycling reaction, comprising the steps of:

(1) contacting the target nucleic acid with the polypeptide of claim 1, and/or the composition of claim 10 or claims 11; and

(2) incubating the target nucleic acid with the polypeptide and/or composition under thermocycling reaction conditions which allow amplification of the target nucleic acid.

21. A host cell transformed with the vector of claim 15.

22. A kit comprising the composition of claim 10 or claim 11, together with packaging materials therefor.

24. A kit comprising the vector of claim 15, together with packaging materials therefor.

25. A kit comprising the host cell of claim 16 or claim 21, together with packaging materials therefor.

8. (canceled)

13. (canceled)

14. (canceled)

19. (canceled)

20. (canceled)

23. A kit comprising the nucleic acid of 12, together with packaging materials therefor.