Thermus thermophilus nucleic acid polymerases
AssigneeApplera Corporation
InventorsJames E. Rozzelle, Elena V. Bolchakova
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Abstract
The invention provides novel nucleic acid polymerases from strains GK24 and RQ-1 of Thermus thermophilus, and nucleic acids encoding those polymerases, as well as methods for using the polymerases and nucleic acids.
Claims55
An isolated nucleic acid encoding a Thermus thermophilus strain RQ-1 (DSM catalog number 9247) nucleic acid polymerase.
An isolated nucleic acid encoding a nucleic acid polymerase comprising any one of amino acid sequences SEQ ID NO:13-24.
An isolated nucleic acid encoding a derivative nucleic acid polymerase comprising any one of amino acid sequences SEQ ID NO:13-15 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:13-15 having a mutation that reduces discrimination against dideoxynucleotide triphosphates.
An isolated nucleic acid encoding a nucleic polymerase comprising any one of SEQ ID NO:1-12.
An isolated nucleic acid comprising a nucleotide sequence complementary to any one of SEQ ID NO:1-12.
A vector comprising an isolated nucleic acid encoding a Thermus thermophilus strain RQ-1 (DSM catalog number 9247) nucleic acid polymerase.
A vector comprising an isolated nucleic acid encoding a nucleic acid polymerase comprising any one of amino acid sequences SEQ ID NO:13-24.
A vector comprising an isolated nucleic acid encoding a derivative nucleic acid polymerase comprising any one of amino acid sequences SEQ ID NO:13-15 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:13-15 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:1-12.
An expression vector comprising a promoter operably linked to an isolated nucleic acid encoding a Thermus thermophilus strain RQ-1 (DSM catalog number 9247) nucleic acid polymerase.
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:13-24.
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:13-15 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:13-15 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:1-12.
A host cell comprising an isolated nucleic acid encoding a Thermus thermophilus strain RQ-1 (DSM catalog number 9247) nucleic acid polymerase.
A host cell comprising an isolated nucleic acid encoding a nucleic acid polymerase comprising any one of amino acid sequences SEQ ID NO:13-24.
A host cell comprising an isolated nucleic acid encoding a derivative nucleic acid polymerase comprising any one of amino acid sequences SEQ ID NO:13-15 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:13-15 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:1-12.
An isolated nucleic acid polymerase from Thermus thermophilus strain RQ-1 (DSM catalog number 9247).
An isolated nucleic acid polymerase comprising any one of amino acid sequences SEQ ID NO:13-24.
An isolated nucleic acid polymerase comprising any one of amino acid sequences SEQ ID NO:13-15 having a mutation that decreases 5-3′ exonuclease activity.
An isolated nucleic acid polymerase comprising any one of amino acid sequences SEQ ID NO:13-15 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:13-24.
A method of making a nucleic acid polymerase comprising any one of SEQ ID NO:13-24 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:13-24 operably linked to a promoter.
A method of synthesizing DNA comprising contacting a polypeptide comprising any one of SEQ ID NO:13-24 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:13-24 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:13-24 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.