PURIFIED THERMOSTABLE PYROCOCCUS FURIOSUS DNA POLYMERASE I

• US 20060040370A1
• Filed: 06/19/2002
• Published: 02/23/2006
• Est. Priority Date: 12/03/1990
• Status: Active Grant

• Current Assignee: Agilent Technologies, Inc.
• Original Assignee: Stratagene California (Agilent Technologies, Inc.)
• Inventors: Mathur, Eric J.
• Granted Patent: US 7,045,328 B2
• Time in Patent Office: Days
• 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: C12Y 207/07007 DNA-directed DNA polymerase (2.7.7.7), i.e. DNA replicase

Abstract

Purified thermostable Pyrococcus furiosus DNA polymerase that migrates on a non-denaturing polyacrylamide gel faster than phosphorylase B and Taq polymerase and more slowly than bovine serum albumin and has an estimated molecular weight of 90,000-93,000 daltons when compared with a Taq polymerase standard assigned a molecular weight of 94,000 daltons.

Claims

1-11. -11. (canceled)

12. An isolated polynucleotide encoding a thermostable DNA polymerase, wherein the isolated polynucleotide is capable of hybridizing to a polynucleotide complementary to a polynucleotide that encodes amino acids 1 to 775 of SEQ ID NO:

1 under hybridization conditions comprising incubating at 42°

C. in a hybridization buffer comprising 0.9 M NaCl and 50% formamide, and under wash conditions comprising incubating at 68°

C. in a wash buffer comprising 0.015 M NaCl and 0.5% sodium dodecyl sulfate.

13. The isolated polynucleotide of claim 12, wherein the thermostable DNA polymerase is a fusion protein.

14. A vector comprising the isolated polynucleotide of claim 12.

15. A host cell comprising the isolated polynucleotide of claim 12.

16. A method of preparing a recombinant thermostable DNA polymerase, comprising culturing the host cell of claim 15 under conditions suitable for the expression of the thermostable DNA polymerase.

17-30. -30. (canceled)