Thermus egertssonii DNA polymerases

• US 8,183,023 B2
• Filed: 10/09/2007
• Issued: 05/22/2012
• Est. Priority Date: 10/09/2006
• Status: Active Grant

• Current Assignee: Qiagen GmbH (Qiagen NV)
• Original Assignee: Qiagen GmbH (Qiagen NV)
• Inventors: Peters, Lars-Erik, Fang, Nan
• Primary Examiner(s): Babic, Christopher M.
• Application Number:
  US12/442,600
• Publication Number:
  US 20090317888A1
• Time in Patent Office: 1,687 Days
• Field of Search: None
• US Class Current: 435/183
• CPC Class Codes: C12N 9/1252 DNA-directed DNA polymerase (2.7.7.7), i.e. DNA replicase
• CPC Class Codes: Y10S 435/975 Kit

Abstract

The present invention relates to a thermophilic polymerase, wherein the DNA polymerase has an in-vitro primer extension rate that is >35 bases/second and faster relative to the primer extension rate of a DNA polymerase comprising amino acid sequences SEQ ID NO: 2 or 4, when measured under identical conditions in a DNA replication assay using primed single strand M13mp18 DNA and an incubation temperature of 60° C. The invention also relates to a vector comprising the polymerase, a host cell comprising the vector. The invention relates to a nucleic acid replication kit comprising the polymerase according to the invention.

Claims

1. An isolated DNA polymerase, wherein the DNA polymerase comprises an amino acid sequence selected from the group consisting of:

SEQ ID NO;

6 and SEQ ID NO;

44.

8. A nucleic acid replication kit comprising a DNA polymerase according to claim 1.

9. The nucleic acid replication kit of claim 8, further comprising a reaction buffer.

10. The nucleic acid replication kit of claim 8, further comprising nucleotides.

11. The nucleic acid replication kit of claim 9, further comprising nucleotides.

12. A nucleic acid replication kit according to claim 8, wherein the kit is selected from the group consisting of a DNA sequencing kit and a DNA amplification kit.

2. An isolated nucleic acid sequence encoding the amino acid sequence selected from the group consisting of:

SEQ ID NO;

6 and SEQ ID NO;

44.

3. A vector comprising the nucleic acid sequence of claim 2.

6. The vector of claim 3, wherein the vector further comprises:

a) a promoter element operably linked to the nucleic acid sequence of claim 2;

b) a ribosome binding site;

c) a selectable metabolic marker gene;

d) an origin of replication functional in a host cell and;

optionally,

e) 3′

-non-translated sequence elements enhancing the translation of the nucleic acid sequence transcript encoding the DNA polymerase.

4. An isolated nucleic acid sequence comprising a sequence selected from the group consisting of:

SEQ ID NO;

5 and SEQ ID NO;

43.

5. A vector comprising the nucleic acid sequence of claim 4.

7. The vector of claim 5, wherein the vector further comprises:

a) a promoter element operably linked to the nucleic acid sequence of claim 4;

b) a ribosome binding site;

c) a selectable metabolic marker gene;

d) an origin of replication functional in a host cell and;

optionally,

e) 3′

-non-translated sequence elements enhancing the translation of the nucleic acid sequence transcript encoding the DNA polymerase.