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

doi:10.3389/fbioe.2020.553474

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 journal › Article › peer-review

9 Scopus citations

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 language	English
Article number	553474
Journal	Frontiers in Bioengineering and Biotechnology
Volume	8
DOIs	https://doi.org/10.3389/fbioe.2020.553474
State	Published - Jan 14 2021

Keywords

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

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10.3389/fbioe.2020.553474

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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.",

keywords = "RNA detection, RT-LAMP, RT-PCR, Taq DNA polymerase, diagnostics, polymerase chain reaction with reverse transcription",

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AU - Zhang, Zhian

AU - Kermekchiev, Milko B.

PY - 2021/1/14

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AB - 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.

KW - RNA detection

KW - RT-LAMP

KW - RT-PCR

KW - Taq DNA polymerase

KW - diagnostics

KW - polymerase chain reaction with reverse transcription

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M3 - Article

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AN - SCOPUS:85102117361

SN - 2296-4185

VL - 8

JO - Frontiers in Bioengineering and Biotechnology

JF - Frontiers in Bioengineering and Biotechnology

M1 - 553474

ER -

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

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