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

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 abil...

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Autores principales: Barnes, Wayne M., Zhang, Zhian, Kermekchiev, Milko B.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7841393/
https://www.ncbi.nlm.nih.gov/pubmed/33520948
http://dx.doi.org/10.3389/fbioe.2020.553474
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author Barnes, Wayne M.
Zhang, Zhian
Kermekchiev, Milko B.
author_facet Barnes, Wayne M.
Zhang, Zhian
Kermekchiev, Milko B.
author_sort Barnes, Wayne M.
collection PubMed
description 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.
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spelling pubmed-78413932021-01-29 A Single Amino Acid Change to Taq DNA Polymerase Enables Faster PCR, Reverse Transcription and Strand-Displacement Barnes, Wayne M. Zhang, Zhian Kermekchiev, Milko B. Front Bioeng Biotechnol Bioengineering and Biotechnology 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. Frontiers Media S.A. 2021-01-14 /pmc/articles/PMC7841393/ /pubmed/33520948 http://dx.doi.org/10.3389/fbioe.2020.553474 Text en Copyright © 2021 Barnes, Zhang and Kermekchiev. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Bioengineering and Biotechnology
Barnes, Wayne M.
Zhang, Zhian
Kermekchiev, Milko B.
A Single Amino Acid Change to Taq DNA Polymerase Enables Faster PCR, Reverse Transcription and Strand-Displacement
title A Single Amino Acid Change to Taq DNA Polymerase Enables Faster PCR, Reverse Transcription and Strand-Displacement
title_full A Single Amino Acid Change to Taq DNA Polymerase Enables Faster PCR, Reverse Transcription and Strand-Displacement
title_fullStr A Single Amino Acid Change to Taq DNA Polymerase Enables Faster PCR, Reverse Transcription and Strand-Displacement
title_full_unstemmed A Single Amino Acid Change to Taq DNA Polymerase Enables Faster PCR, Reverse Transcription and Strand-Displacement
title_short A Single Amino Acid Change to Taq DNA Polymerase Enables Faster PCR, Reverse Transcription and Strand-Displacement
title_sort single amino acid change to taq dna polymerase enables faster pcr, reverse transcription and strand-displacement
topic Bioengineering and Biotechnology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7841393/
https://www.ncbi.nlm.nih.gov/pubmed/33520948
http://dx.doi.org/10.3389/fbioe.2020.553474
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