Thermostable type-A DNA polymerase mutants with increased polymerization rate and resistance to inhibitors

摘要

The present invention provides mutants of DNA polymerases having an increased rate of incorporation of nucleotides into nucleic acids undergoing polymerization and having an enhanced resistance to inhibitors of DNA polymerase activity. The mutant polymerases are well suited for fast PCR applications, for PCR amplification of targets in samples that contain inhibitors of wild-type polymerases, and for fast PCR amplification of samples containing DNA polymerase inhibitors. In exemplary embodiments, the mutants are mutants of Taq DNA polymerase.

主权项

1. A mutant thermostable Type-A DNA polymerase consisting of or comprising:
a first mutation at residue 507 of wild-type Taq DNA polymerase, which has the sequence of SEQ ID NO:4, or at a residue corresponding to residue 507 of wild-type Taq DNA polymerase in another thermostable Type-A DNA polymerase; and at least one additional mutation at a residue selected from 59, 155, 245, 375, 508, 734, and 749 of wild-type Taq DNA polymerase, or at a corresponding residue in another thermostable Type-A DNA polymerase, wherein the combination of mutations provides a mutant polymerase that possesses a faster polymerization rate and a higher resistance to polymerization activity inhibitors than the wild-type DNA polymerase from which it is derived.