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Optimization of production of recombinant gamma-tubulin in bacteria

Zhou, Jingkai LU and Alvarado Kristensson, Maria LU (2021) In MethodsX 8. p.1-8
Abstract
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 lysates using affinity tags.
 This protocol describes a method that purifies soluble GST-TUBG1 from bacteria.
 Of the three tested induction conditions, the highest yield... (More)
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 lysates using affinity tags.
 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 [1], the technique described here retrieves larger amounts of recombinant γ-tubulin from small-scale expression cultures.
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author
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organization
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type
Contribution to journal
publication status
published
subject
in
MethodsX
volume
8
article number
101517
pages
1 - 8
publisher
Elsevier
external identifiers
  • scopus:85114993353
  • pmid:34754788
ISSN
2215-0161
DOI
10.1016/j.mex.2021.101517
language
English
LU publication?
yes
id
f2c8238b-375d-43c4-9c72-7ef02edc7d73
date added to LUP
2021-09-16 19:30:50
date last changed
2022-05-05 03:41:56
@article{f2c8238b-375d-43c4-9c72-7ef02edc7d73,
  abstract     = {{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 lysates using affinity tags.<br/>	This protocol describes a method that purifies soluble GST-TUBG1 from bacteria. <br/>	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.<br/>	In comparison with other methodologies [1], the technique described here retrieves larger amounts of recombinant γ-tubulin from small-scale expression cultures.<br/>}},
  author       = {{Zhou, Jingkai and Alvarado Kristensson, Maria}},
  issn         = {{2215-0161}},
  language     = {{eng}},
  pages        = {{1--8}},
  publisher    = {{Elsevier}},
  series       = {{MethodsX}},
  title        = {{Optimization of production of recombinant gamma-tubulin in bacteria}},
  url          = {{http://dx.doi.org/10.1016/j.mex.2021.101517}},
  doi          = {{10.1016/j.mex.2021.101517}},
  volume       = {{8}},
  year         = {{2021}},
}