Efficient and high fidelity incorporation of dye-terminators by a novel archaeal DNA polymerase mutant.

@article{Arezi2002EfficientAH,
  title={Efficient and high fidelity incorporation of dye-terminators by a novel archaeal DNA polymerase mutant.},
  author={Bahram Arezi and Connie J. Hansen and Holly H. Hogrefe},
  journal={Journal of molecular biology},
  year={2002},
  volume={322 4},
  pages={719-29}
}
We examined the molecular basis of ddNTP selectivity in archaeal family B DNA polymerases by randomly mutagenizing the gene encoding Thermococcus sp. JDF-3 DNA polymerase and screening mutant libraries for improved ddNTP incorporation. We identified two mutations, P410L and A485T, that improved ddNTP uptake, suggesting the contribution of P410 and A485 to ddNTP/dNTP selectivity in archaeal DNA polymerases. The importance of A485 was identified previously in mutagenesis studies employing Pfu… CONTINUE READING