On the fidelity of DNA replication. Isolation of high fidelity DNA polymerase-primase complexes by immunoaffinity chromatography.

preliminary expression of high fidelity pfu dna polymerase encoding fragment

High fidelity DNA synthesis by the Thermus aquaticus DNA polymerase

We demonstrate that despite lacking a 3'----5' proofreading exonuclease, the Thermus aquaticus (Taq) DNA polymerase can catalyze highly accurate DNA synthesis in vitro. Under defined reaction conditions, the error rate per nucleotide polymerized at 70 degrees C can be as low as 10(-5) for base substitution errors and 10(-6) for frameshift errors. The frequency of mutations produced during a sin...

Calcium-driven DNA synthesis by a high-fidelity DNA polymerase

Divalent metal ions, usually Mg2+, are required for both DNA synthesis and proofreading functions by DNA polymerases (DNA Pol). Although used as a non-reactive cofactor substitute for binding and crystallographic studies, Ca2+ supports DNA polymerization by only one DNA Pol, Dpo4. Here, we explore whether Ca2+-driven catalysis might apply to high-fidelity (HiFi) family B DNA Pols. The consequen...

Kunkel on DNA Replication Fidelity

Background: Caenorhabditis elegans is an important model for the study of DNA damage and repair related processes such as aging, neurodegeneration, and carcinogenesis. However, DNA repair is poorly characterized in this organism. We adapted a quantitative polymerase chain reaction assay to characterize repair of DNA damage induced by ultraviolet type C (UVC) radiation in C. elegans, and then te...

On the Fidelity of DNA Replication

Homogeneous DNA polymerase (“reverse transcriptase”) from avian myeloblastosis virus was assayed for exodeoxyribonuclease activity. The substrates were defined template. initiator complexes in which different radioactive nucleotides were present at the 3’-OH termini of the initiator. Even when the number of molecules of enzyme was equal to the number of initiator termini, there was no significa...