Total chemical synthesis of a thermostable enzyme capable of polymerase chain reaction

Polymerase chain reaction (PCR) has been a defining tool in modern biology. Towards realizing mirror-image PCR, we have designed and chemically synthesized a mutant version of the 352-residue thermostable Sulfolobus solfataricus P2 DNA polymerase IV with l-amino acids and tested its PCR activity bio...

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Autores principales: Xu, Weiliang, Jiang, Wenjun, Wang, Jiaxing, Yu, Linping, Chen, Ji, Liu, Xianyu, Liu, Lei, Zhu, Ting F
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group 2017
Materias:
Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5335361/
https://www.ncbi.nlm.nih.gov/pubmed/28265464
http://dx.doi.org/10.1038/celldisc.2017.8
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author Xu, Weiliang
Jiang, Wenjun
Wang, Jiaxing
Yu, Linping
Chen, Ji
Liu, Xianyu
Liu, Lei
Zhu, Ting F
author_facet Xu, Weiliang
Jiang, Wenjun
Wang, Jiaxing
Yu, Linping
Chen, Ji
Liu, Xianyu
Liu, Lei
Zhu, Ting F
author_sort Xu, Weiliang
collection PubMed
description Polymerase chain reaction (PCR) has been a defining tool in modern biology. Towards realizing mirror-image PCR, we have designed and chemically synthesized a mutant version of the 352-residue thermostable Sulfolobus solfataricus P2 DNA polymerase IV with l-amino acids and tested its PCR activity biochemically. To the best of our knowledge, this enzyme is the largest chemically synthesized protein reported to date. We show that with optimization of PCR conditions, the fully synthetic polymerase is capable of amplifying template sequences of up to 1.5 kb. The establishment of this synthetic route for chemically synthesizing DNA polymerase IV is a stepping stone towards building a d-enzyme system for mirror-image PCR, which may open up an avenue for the creation of many mirror-image molecular tools such as mirror-image systematic evolution of ligands by exponential enrichment.
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spelling pubmed-53353612017-03-06 Total chemical synthesis of a thermostable enzyme capable of polymerase chain reaction Xu, Weiliang Jiang, Wenjun Wang, Jiaxing Yu, Linping Chen, Ji Liu, Xianyu Liu, Lei Zhu, Ting F Cell Discov Article Polymerase chain reaction (PCR) has been a defining tool in modern biology. Towards realizing mirror-image PCR, we have designed and chemically synthesized a mutant version of the 352-residue thermostable Sulfolobus solfataricus P2 DNA polymerase IV with l-amino acids and tested its PCR activity biochemically. To the best of our knowledge, this enzyme is the largest chemically synthesized protein reported to date. We show that with optimization of PCR conditions, the fully synthetic polymerase is capable of amplifying template sequences of up to 1.5 kb. The establishment of this synthetic route for chemically synthesizing DNA polymerase IV is a stepping stone towards building a d-enzyme system for mirror-image PCR, which may open up an avenue for the creation of many mirror-image molecular tools such as mirror-image systematic evolution of ligands by exponential enrichment. Nature Publishing Group 2017-02-28 /pmc/articles/PMC5335361/ /pubmed/28265464 http://dx.doi.org/10.1038/celldisc.2017.8 Text en Copyright © 2017 The Author(s) http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/
spellingShingle Article
Xu, Weiliang
Jiang, Wenjun
Wang, Jiaxing
Yu, Linping
Chen, Ji
Liu, Xianyu
Liu, Lei
Zhu, Ting F
Total chemical synthesis of a thermostable enzyme capable of polymerase chain reaction
title Total chemical synthesis of a thermostable enzyme capable of polymerase chain reaction
title_full Total chemical synthesis of a thermostable enzyme capable of polymerase chain reaction
title_fullStr Total chemical synthesis of a thermostable enzyme capable of polymerase chain reaction
title_full_unstemmed Total chemical synthesis of a thermostable enzyme capable of polymerase chain reaction
title_short Total chemical synthesis of a thermostable enzyme capable of polymerase chain reaction
title_sort total chemical synthesis of a thermostable enzyme capable of polymerase chain reaction
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5335361/
https://www.ncbi.nlm.nih.gov/pubmed/28265464
http://dx.doi.org/10.1038/celldisc.2017.8
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