Improved designs for pET expression plasmids increase protein production yield in Escherichia coli

The pET series of expression plasmids are widely used for recombinant protein production in Escherichia coli. The genetic modules controlling transcription and translation in these plasmids were first described in the 1980s and have not changed since. Herein we report design flaws in these genetic m...

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Autores principales: Shilling, Patrick J., Mirzadeh, Kiavash, Cumming, Alister J., Widesheim, Magnus, Köck, Zoe, Daley, Daniel O.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2020
Materias:
Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7205610/
https://www.ncbi.nlm.nih.gov/pubmed/32382055
http://dx.doi.org/10.1038/s42003-020-0939-8
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author Shilling, Patrick J.
Mirzadeh, Kiavash
Cumming, Alister J.
Widesheim, Magnus
Köck, Zoe
Daley, Daniel O.
author_facet Shilling, Patrick J.
Mirzadeh, Kiavash
Cumming, Alister J.
Widesheim, Magnus
Köck, Zoe
Daley, Daniel O.
author_sort Shilling, Patrick J.
collection PubMed
description The pET series of expression plasmids are widely used for recombinant protein production in Escherichia coli. The genetic modules controlling transcription and translation in these plasmids were first described in the 1980s and have not changed since. Herein we report design flaws in these genetic modules. We present improved designs and demonstrate that, when incorporated into pET28a, they support increases in protein production. The improved designs are applicable to most of the 103 vectors in the pET series and can be easily implemented.
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spelling pubmed-72056102020-05-14 Improved designs for pET expression plasmids increase protein production yield in Escherichia coli Shilling, Patrick J. Mirzadeh, Kiavash Cumming, Alister J. Widesheim, Magnus Köck, Zoe Daley, Daniel O. Commun Biol Article The pET series of expression plasmids are widely used for recombinant protein production in Escherichia coli. The genetic modules controlling transcription and translation in these plasmids were first described in the 1980s and have not changed since. Herein we report design flaws in these genetic modules. We present improved designs and demonstrate that, when incorporated into pET28a, they support increases in protein production. The improved designs are applicable to most of the 103 vectors in the pET series and can be easily implemented. Nature Publishing Group UK 2020-05-07 /pmc/articles/PMC7205610/ /pubmed/32382055 http://dx.doi.org/10.1038/s42003-020-0939-8 Text en © The Author(s) 2020 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Article
Shilling, Patrick J.
Mirzadeh, Kiavash
Cumming, Alister J.
Widesheim, Magnus
Köck, Zoe
Daley, Daniel O.
Improved designs for pET expression plasmids increase protein production yield in Escherichia coli
title Improved designs for pET expression plasmids increase protein production yield in Escherichia coli
title_full Improved designs for pET expression plasmids increase protein production yield in Escherichia coli
title_fullStr Improved designs for pET expression plasmids increase protein production yield in Escherichia coli
title_full_unstemmed Improved designs for pET expression plasmids increase protein production yield in Escherichia coli
title_short Improved designs for pET expression plasmids increase protein production yield in Escherichia coli
title_sort improved designs for pet expression plasmids increase protein production yield in escherichia coli
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7205610/
https://www.ncbi.nlm.nih.gov/pubmed/32382055
http://dx.doi.org/10.1038/s42003-020-0939-8
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