Mutant RB69 DNA polymerase

Patent metadata

Field	Value
Publication number	US-9399767-B2
Application number	US-201414191997-A
Country	US
Kind code	B2
Filing date	Feb 27, 2014
Priority date	Jun 5, 2009
Publication date	Jul 26, 2016
Grant date	Jul 26, 2016

Abstract

Provided herein are mutant DNA-dependent polymerases which are derived from, or otherwise related to, wild type RB69 DNA polymerase. These mutant polymerases are capable of selectively binding labeled nucleotides. These mutant polymerases are also capable of incorporating a variety of naturally occurring and modified nucleotides, including, for example, terminator nucleotides.

First claim

What is claimed:

1. A system comprising a mutant DNA polymerase bound to a DNA template and a primer, and a nucleotide transiently-bound to the mutant DNA polymerase, wherein the mutant DNA polymerase comprises the amino acid of SEQ ID NO:2.

2. The system of claim 1 , further comprising a metal cation that inhibits incorporation of the transiently-bound nucleotide.

3. The system of claim 2 , wherein the metal cation is selected from a group consisting of calcium, scandium, titanium, vanadium, chromium, iron, cobalt, nickel, copper, zinc, gallium, germanium, arsenic, and selenium.

4. The system of claim 1 , wherein the transiently-bound nucleotide is linked to a report moiet.

5. The system of claim 1 , wherein the mutant DNA polymerase further comprises a reporter moiety.

6. The system of claim 5 , wherein the reporter moiety comprises an energy transfer donor moiety.

7. The system of claim 5 , wherein the reporter moiety comprises a fluorescent dye or a nanoparticle.

8. The system of claim 1 , wherein the transiently-bound nucleotide comprises a reversible terminator nucleotide.

9. The system of claim 8 , wherein the reversible terminator nucleotide comprises an azidomethyl blocking group on the 3′ sugar position.

Assignees

• Life Technologies Corp
• Lift Tech Corp

Inventors

• Peris Marian
• Phelan Michael
• Rosenblum Barnett
• Hendricks Stephen

Classifications

• C12Q1/6869

  Methods for sequencing · CPC title

• C12Q1/6872

  involving mass spectrometry · CPC title

• C12Q1/6827

  for detection of mutation or polymorphism · CPC title

• C12Q1/6804

  Nucleic acid analysis using immunogens (immunoassay G01N33/53) · CPC title

• G01N2500/00

  Screening for compounds of potential therapeutic value · CPC title