Thermus scotoductus nucleic acid polymerases

• US 9,587,264 B2
• Filed: 10/21/2015
• Issued: 03/07/2017
• Est. Priority Date: 09/14/2001
• Status: Active Grant

• Current Assignee: Applied Biosystems LLC (Thermo Fisher Scientific Incorporated)
• Original Assignee: Applied Biosystems LLC (Thermo Fisher Scientific Incorporated)
• Inventors: Bolchakova, Elena, Rozzelle, James
• Primary Examiner(s): HUTSON, RICHARD G
• Application Number:
  US14/919,679
• Publication Number:
  US 20160115512A1
• Time in Patent Office: 503 Days
• Field of Search: None
• US Class Current: 1/1
• CPC Class Codes: C12N 9/1252 DNA-directed DNA polymerase (2.7.7.7), i.e. DNA replicase
• CPC Class Codes: C12P 19/34 Polynucleotides, e.g. nucleic acids, oligoribonucleotides
• CPC Class Codes: C12Q 1/6806 Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay C12Q1/6804 takes precedence
• CPC Class Codes: C12Q 1/686 Polymerase chain reaction [PCR]
• CPC Class Codes: C12Q 1/6869 Methods for sequencing
• CPC Class Codes: C12Y 207/07007 DNA-directed DNA polymerase (2.7.7.7), i.e. DNA replicase
• CPC Class Codes: Y02P 20/52 using catalysts, e.g. selective catalysts

Abstract

The invention provides nucleic acids and polypeptides for a nucleic acid polymerase from a thermophilic organism, Thermus scotoductus. The invention also provides methods for using these nucleic acids and polypeptides.

Claims

1. A method of synthesizing DNA comprising contacting a polypeptide having at least 95% sequence identity to SEQ ID NO:

14, wherein said polypeptide comprises an Asp at a position corresponding to amino acid 46 and an Arg at a position corresponding to amino acid 250 of SEQ ID NO;

13, with a DNA under conditions sufficient to permit polymerization of DNA.

2. A method for thermocyclic amplification of nucleic acid comprising:

(a) contacting a nucleic acid with a thermostable polypeptide having at least 95% sequence identity to SEQ ID NO;

14, wherein said polypeptide comprises an Asp at a position corresponding to amino acid 46 and an Arg at a position corresponding to amino acid 250 of SEQ ID NO;

13 under conditions suitable for amplification of said nucleic acid; and

(b) amplifying the nucleic acid.

3. The method of claim 2 wherein the thermocyclic amplification of the nucleic acid includes cycles of denaturation, primer annealing and primer extension.

4. The method of claim 2 wherein the thermocyclic amplification of the nucleic acid is performed by Strand Displacement Amplification.

5. The method of claim 2 wherein thermocyclic amplification of the nucleic acid is performed by Polymerase Chain Reaction.

6. A method of primer extension comprising contacting a polypeptide having at least 95% sequence identity to SEQ ID NO:

14, wherein said polypeptide comprises an Asp at a position corresponding to amino acid 46 and an Arg at a position corresponding to amino acid 250 of SEQ ID NO;

13 with a primer and a nucleic acid that is complementary to the primer under conditions sufficient to permit polymerization of DNA.

7. The method of claim 6 wherein the nucleic acid is DNA.

8. The method of claim 6 wherein the primer extension is done to sequence the nucleic acid.

9. The method of claim 6 wherein the primer extension is done to amplify the nucleic acid.