Cargando…
Optimization of production of recombinant gamma-tubulin in bacteria
Production of a protein of interest in bacteria and its purification from bacterial lysates are valuable tools for the purification of larger amounts of recombinant proteins. The low cost of culturing, and the rapid cell growth of bacteria make this host a good choice for protein production, but the...
Autores principales: | , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier
2021
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8563660/ https://www.ncbi.nlm.nih.gov/pubmed/34754788 http://dx.doi.org/10.1016/j.mex.2021.101517 |
_version_ | 1784593451532156928 |
---|---|
author | Zhou, Jingkai Alvarado-Kristensson, Maria |
author_facet | Zhou, Jingkai Alvarado-Kristensson, Maria |
author_sort | Zhou, Jingkai |
collection | PubMed |
description | Production of a protein of interest in bacteria and its purification from bacterial lysates are valuable tools for the purification of larger amounts of recombinant proteins. The low cost of culturing, and the rapid cell growth of bacteria make this host a good choice for protein production, but the folding and function of the purified protein might be altered due to the production of a eukaryotic protein in a prokaryotic host. Here, we provide a purification method for the purification of gamma (γ)-tubulin (TUBG) from soluble fractions of Escherichia (E.) coli • This protocol describes a method that purifies soluble GST-TUBG1 from bacteria. • Of the three tested induction conditions, the highest yield of recombinant GST-TUBG1 was obtained after the induction of E. coli with isopropyl-D-1-thiogalactopyranoside (IPTG) for 1 h at 37 °C followed by overnight incubation at room temperature. • In comparison with other methodologies (Hoog et al., 2011), the technique described here retrieves larger amounts of recombinant TUBG1 from small-scale expression cultures. |
format | Online Article Text |
id | pubmed-8563660 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
spelling | pubmed-85636602021-11-08 Optimization of production of recombinant gamma-tubulin in bacteria Zhou, Jingkai Alvarado-Kristensson, Maria MethodsX Method Article Production of a protein of interest in bacteria and its purification from bacterial lysates are valuable tools for the purification of larger amounts of recombinant proteins. The low cost of culturing, and the rapid cell growth of bacteria make this host a good choice for protein production, but the folding and function of the purified protein might be altered due to the production of a eukaryotic protein in a prokaryotic host. Here, we provide a purification method for the purification of gamma (γ)-tubulin (TUBG) from soluble fractions of Escherichia (E.) coli • This protocol describes a method that purifies soluble GST-TUBG1 from bacteria. • Of the three tested induction conditions, the highest yield of recombinant GST-TUBG1 was obtained after the induction of E. coli with isopropyl-D-1-thiogalactopyranoside (IPTG) for 1 h at 37 °C followed by overnight incubation at room temperature. • In comparison with other methodologies (Hoog et al., 2011), the technique described here retrieves larger amounts of recombinant TUBG1 from small-scale expression cultures. Elsevier 2021-09-16 /pmc/articles/PMC8563660/ /pubmed/34754788 http://dx.doi.org/10.1016/j.mex.2021.101517 Text en © 2021 The Authors. Published by Elsevier B.V. https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). |
spellingShingle | Method Article Zhou, Jingkai Alvarado-Kristensson, Maria Optimization of production of recombinant gamma-tubulin in bacteria |
title | Optimization of production of recombinant gamma-tubulin in bacteria |
title_full | Optimization of production of recombinant gamma-tubulin in bacteria |
title_fullStr | Optimization of production of recombinant gamma-tubulin in bacteria |
title_full_unstemmed | Optimization of production of recombinant gamma-tubulin in bacteria |
title_short | Optimization of production of recombinant gamma-tubulin in bacteria |
title_sort | optimization of production of recombinant gamma-tubulin in bacteria |
topic | Method Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8563660/ https://www.ncbi.nlm.nih.gov/pubmed/34754788 http://dx.doi.org/10.1016/j.mex.2021.101517 |
work_keys_str_mv | AT zhoujingkai optimizationofproductionofrecombinantgammatubulininbacteria AT alvaradokristenssonmaria optimizationofproductionofrecombinantgammatubulininbacteria |