Frontiers in Bioengineering and Biotechnology (Jan 2021)

A Single Amino Acid Change to Taq DNA Polymerase Enables Faster PCR, Reverse Transcription and Strand-Displacement

  • Wayne M. Barnes,
  • Wayne M. Barnes,
  • Zhian Zhang,
  • Milko B. Kermekchiev

DOI
https://doi.org/10.3389/fbioe.2020.553474
Journal volume & issue
Vol. 8

Abstract

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A change of an aspartic acid to asparagine of Taq (Thermus aquaticus) DNA polymerase is a gain of function mutation that supports faster PCR: the extension times for PCR amplification can be 2–3 times shorter. Surprising results from negative controls led to the discovery of strand-displacement ability and reverse transcriptase activity of Taq D732N DNA polymerase. We demonstrate that the mutant enzyme can, by itself, catalyze RT-PCR, and RT-LAMP assays. Residue 732 is on the surface of the enzyme, not near the active site.

Keywords