Thermus brockianus nucleic acid polymerases
AssigneeApplera Corporation
InventorsJames E. Rozzelle, Elena V. Bolchakova
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Abstract
The invention provides nucleic acids and polypeptides for nucleic acid polymerases from a thermophilic organism, Thermus brockianus. The invention also provides methods for using these nucleic acids and polypeptides.
Claims54
An isolated nucleic acid encoding a nucleic acid polymerase comprising any one of amino acid sequences SEQ ID NO:9-16.
An isolated nucleic acid encoding a derivative nucleic acid polymerase comprising any one of amino acid sequences SEQ ID NO:9-10 having a mutation that decreases 5-3′ exonuclease activity.
An isolated nucleic acid encoding a derivative nucleic acid polymerase comprising any one of amino acid sequences SEQ ID NO:
- 9-10 having a mutation that reduces discrimination against dideoxynucleotide triphosphates.
An isolated nucleic acid encoding a nucleic polymerase comprising any one of SEQ ID NO:1-8.
An isolated nucleic acid comprising a nucleotide sequence complementary to any one of SEQ ID NO:1-8.
An isolated nucleic acid encoding a nucleic acid polymerase from Thermus brockianus comprising amino acid sequence with at least 96% identity to any one of SEQ ID NO:9-16.
A vector comprising an isolated nucleic acid encoding a nucleic acid polymerase comprising any one of amino acid sequences SEQ ID NO:9-16.
A vector comprising an isolated nucleic acid encoding a derivative nucleic acid polymerase comprising any one of amino acid sequences SEQ ID NO:9-10 having a mutation that decreases 5-3′ exonuclease activity.
A vector comprising an isolated nucleic acid encoding a derivative nucleic acid polymerase comprising any one of amino acid sequences SEQ ID NO:
- 9-10 having a mutation that reduces discrimination against dideoxynucleotide triphosphates.
A vector comprising an isolated nucleic acid encoding a nucleic polymerase comprising any one of SEQ ID NO:1-8.
A vector comprising an isolated nucleic acid comprising a nucleotide sequence complementary to any one of SEQ ID NO:1-8.
A vector comprising an isolated nucleic acid encoding a nucleic acid polymerase from Thermus brockianus comprising amino acid sequence with at least 96% identity to any one of SEQ ID NO:9-16.
An expression vector comprising a promoter operably linked to an isolated nucleic acid encoding a nucleic acid polymerase comprising any one of amino acid sequences SEQ ID NO:9-16.
An expression vector comprising a promoter operably linked to an isolated nucleic acid encoding a derivative nucleic acid polymerase comprising any one of amino acid sequences SEQ ID NO:9-10 having a mutation that decreases 5-3′ exonuclease activity.
An expression vector comprising a promoter operably linked to an isolated nucleic acid encoding a derivative nucleic acid polymerase comprising any one of amino acid sequences SEQ ID NO:9-10 having a mutation that reduces discrimination against dideoxynucleotide triphosphates.
An expression vector comprising a promoter operably linked to an isolated nucleic acid encoding a nucleic polymerase comprising any one of SEQ ID NO:1-8.
An expression vector comprising a promoter operably linked to an isolated nucleic acid comprising a nucleotide sequence complementary to any one of SEQ ID NO:1-8.
An expression vector comprising a promoter operably linked to an isolated nucleic acid encoding a nucleic acid polymerase from Thermus brockianus comprising amino acid sequence with at least 96% identity to any one of SEQ ID NO:9-16.
A host cell comprising an isolated nucleic acid encoding a nucleic acid polymerase comprising any one of amino acid sequences SEQ ID NO:9-16.
A host cell comprising an isolated nucleic acid encoding a derivative nucleic acid polymerase comprising any one of amino acid sequences SEQ ID NO:9-10 having a mutation that decreases 5-3′ exonuclease activity.
A host cell comprising an isolated nucleic acid encoding a derivative nucleic acid polymerase comprising any one of amino acid sequences SEQ ID NO:
- 9-10 having a mutation that reduces discrimination against dideoxynucleotide triphosphates.
A host cell comprising an isolated nucleic acid encoding a nucleic polymerase comprising any one of SEQ ID NO:1-8.
A host cell comprising an isolated nucleic acid encoding a nucleic acid polymerase from Thermus brockianus comprising amino acid sequence with at least 96% identity to any one of SEQ ID NO:9-16.
An isolated nucleic acid polymerase comprising any one of amino acid sequences SEQ ID NO:9-16.
An isolated nucleic acid polymerase comprising any one of amino acid sequences SEQ ID NO:9-49.
An isolated nucleic acid polymerase comprising any one of amino acid sequences SEQ ID NO:9-10 having a mutation that decreases 5-3′ exonuclease activity.
An isolated nucleic acid polymerase comprising any one of amino acid sequences SEQ ID NO:9-10 having a mutation that reduces discrimination against dideoxynucleotide triphosphates.
A kit comprising a container containing a nucleic acid polymerase, wherein the nucleic acid polymerase comprises any one of amino acid sequences SEQ ID NO:9-49.
A kit comprising a container containing a nucleic acid polymerase, wherein the nucleic acid polymerase comprises any one of amino acid sequences SEQ ID NO:9-16.
A method of making a nucleic acid polymerase comprising any one of SEQ ID NO:9-16 that comprises, incubating under conditions sufficient for RNA transcription and translation a host cell comprising a nucleic acid encoding a polypeptide comprising any one of SEQ ID NO:9-16 operably linked to a promoter.
A method of synthesizing DNA comprising contacting a polypeptide comprising any one of SEQ ID NO:9-16 with a DNA under conditions sufficient to permit polymerization of DNA.
A method of synthesizing DNA comprising contacting a polypeptide comprising any one of SEQ ID NO:9-49 with a DNA under conditions sufficient to permit polymerization of DNA.
A method for thermocyclic amplification of nucleic acid comprising:
- (a) contacting a nucleic acid with a thermostable polypeptide having any one of SEQ ID NO:9-49 under conditions suitable for amplification of said nucleic acid;
- and
- (b) amplifying the nucleic acid.
A method of primer extension comprising contacting a polypeptide comprising any one of SEQ ID NO:9-16 with a primer and a nucleic acid that is complementary to the primer under conditions sufficient to permit polymerization of DNA.
Dependencies are inferred from the claim text.