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Abstract

This invention provides DNA polymerases with mutations in the charge-switch nucleotide interaction region that increase activity for charge-switch nucleotides. Such polymerases can be generated by introducing mutations in specific residues which are identified as being in the appropriate region through structural models, by homology to polymerases with known structures, or experimental analysis. In some embodiments, the mutant DNA polymerases have additional mutations that decrease activity for non-charge-switch nucleotides and mutations that decrease exonuclease activity. In another aspect, the invention provides methods of sequencing a target nucleic acid with the above described mutated DNA polymerases. In yet another aspect, the invention provides methods of generating polypeptides having charge-switch nucleotide polymerase activity by introducing 'random' mutations and selecting those mutated polypeptides that encode polypeptides having charge-switch nucleotide activity

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First Claim