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An Escherichia coli strain for expression of the connexin45 carboxyl terminus attached to the 4th transmembrane domain
A major problem for structural characterization of membrane proteins, such as connexins, by nuclear magnetic resonance (NMR) occurs at the initial step of the process, the production of sufficient amounts of protein. This occurs because proteins must be expressed in minimal based media. Here, we des...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Frontiers Media S.A.
2013
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3750199/ https://www.ncbi.nlm.nih.gov/pubmed/23986705 http://dx.doi.org/10.3389/fphar.2013.00106 |
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author | Kopanic, Jennifer L. Al-Mugotir, Mona Zach, Sydney Das, Srustidhar Grosely, Rosslyn Sorgen, Paul L. |
author_facet | Kopanic, Jennifer L. Al-Mugotir, Mona Zach, Sydney Das, Srustidhar Grosely, Rosslyn Sorgen, Paul L. |
author_sort | Kopanic, Jennifer L. |
collection | PubMed |
description | A major problem for structural characterization of membrane proteins, such as connexins, by nuclear magnetic resonance (NMR) occurs at the initial step of the process, the production of sufficient amounts of protein. This occurs because proteins must be expressed in minimal based media. Here, we describe an expression system for membrane proteins that significantly improves yield by addressing two common problems, cell toxicity caused by protein translation and codon bias between genomes. This work provides researchers with a cost-effective tool for NMR and other biophysical studies, to use when faced with little-to-no expression of eukaryotic membrane proteins in Escherichia coli expression systems. |
format | Online Article Text |
id | pubmed-3750199 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2013 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-37501992013-08-28 An Escherichia coli strain for expression of the connexin45 carboxyl terminus attached to the 4th transmembrane domain Kopanic, Jennifer L. Al-Mugotir, Mona Zach, Sydney Das, Srustidhar Grosely, Rosslyn Sorgen, Paul L. Front Pharmacol Pharmacology A major problem for structural characterization of membrane proteins, such as connexins, by nuclear magnetic resonance (NMR) occurs at the initial step of the process, the production of sufficient amounts of protein. This occurs because proteins must be expressed in minimal based media. Here, we describe an expression system for membrane proteins that significantly improves yield by addressing two common problems, cell toxicity caused by protein translation and codon bias between genomes. This work provides researchers with a cost-effective tool for NMR and other biophysical studies, to use when faced with little-to-no expression of eukaryotic membrane proteins in Escherichia coli expression systems. Frontiers Media S.A. 2013-08-23 /pmc/articles/PMC3750199/ /pubmed/23986705 http://dx.doi.org/10.3389/fphar.2013.00106 Text en Copyright © Kopanic, Al-Mugotir, Zach, Das, Grosely and Sorgen. http://creativecommons.org/licenses/by/3.0/ 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) or licensor 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 | Pharmacology Kopanic, Jennifer L. Al-Mugotir, Mona Zach, Sydney Das, Srustidhar Grosely, Rosslyn Sorgen, Paul L. An Escherichia coli strain for expression of the connexin45 carboxyl terminus attached to the 4th transmembrane domain |
title | An Escherichia coli strain for expression of the connexin45 carboxyl terminus attached to the 4th transmembrane domain |
title_full | An Escherichia coli strain for expression of the connexin45 carboxyl terminus attached to the 4th transmembrane domain |
title_fullStr | An Escherichia coli strain for expression of the connexin45 carboxyl terminus attached to the 4th transmembrane domain |
title_full_unstemmed | An Escherichia coli strain for expression of the connexin45 carboxyl terminus attached to the 4th transmembrane domain |
title_short | An Escherichia coli strain for expression of the connexin45 carboxyl terminus attached to the 4th transmembrane domain |
title_sort | escherichia coli strain for expression of the connexin45 carboxyl terminus attached to the 4th transmembrane domain |
topic | Pharmacology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3750199/ https://www.ncbi.nlm.nih.gov/pubmed/23986705 http://dx.doi.org/10.3389/fphar.2013.00106 |
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