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

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

Research output: Contribution to journalArticlepeer-review

Abstract

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.

Original languageEnglish
Article number553474
JournalFrontiers in Bioengineering and Biotechnology
Volume8
DOIs
StatePublished - Jan 14 2021

Keywords

  • RNA detection
  • RT-LAMP
  • RT-PCR
  • Taq DNA polymerase
  • diagnostics
  • polymerase chain reaction with reverse transcription

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