Patent
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.
1. An isolated mutant nucleic acid polymerase comprising SEQ ID NO: 20.
2. The isolated mutant nucleic acid polymerase of claim 1 , wherein the polymerase exhibits decreased 5-3′ exonuclease activity relative to its corresponding wild type nucleic acid polymerase.
3. The isolated mutant nucleic acid polymerase of claim 1 , wherein the polymerase exhibits reduced discrimination against dideoxynucleotide triphosphates relative to its corresponding wild type nucleic acid polymerase.
4. A kit comprising a container containing the mutant nucleic acid polymerase of claim 1 .
5. The kit of claim 4 further comprising a container containing an unlabeled nucleotide, a labeled nucleotide, a balanced mixture of nucleotides, a chain terminating nucleotide, a nucleotide analog, a buffer solution, a solution containing magnesium, a cloning vector, a restriction endonuclease, a sequencing primer, a solution containing reverse transcriptase, or a DNA or RNA amplification primer.
6. The kit of claim 4 , adapted for performing DNA sequencing, DNA amplification, reverse transcription, RNA amplification or primer extension.
7. The isolated mutant nucleic acid polymerase of claim 1 , wherein the polymerase comprises an amino acid substitution at position 46.
8. The isolated mutant nucleic acid polymerase of claim 7 , wherein the amino acid substitution at position 46 comprises a substitution with Asp.
9. The isolated mutant nucleic acid polymerase of claim 1 , wherein the polymerase is made by incubating a host cell under conditions sufficient for RNA transcription and translation of a nucleic acid encoding the polymerase operably linked to a promoter.