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Title: T7 Expression Systems for Inducible Production of Proteins from Cloned Genes in E. coli

Abstract

Inducible T7 expression systems are capable of producing a wide range of proteins in E. coli. Improvements over common practice include: preventing unintended induction by establishing and maintaining expression strains in non-inducing growth media comprised entirely of purified components instead of complex growth media that may variably induce target proteins on approach to saturation; and expressing many target proteins in parallel by convenient and productive auto-induction in BL21(DE3) and other suitable hosts, instead of IPTG induction. From earliest days, basal expression prevented establishment of inducible strains for producing proteins that are stressful to the host. Newly developed pAL vectors now reduce basal expression to levels where coding sequences for even the most stressful proteins can be maintained and induced. In conclusion, asymmetric ligation allows simple and efficient cloning of individual coding sequences or simultaneous cloning of two or three coding sequences for co expression from a single pAL vector.

Authors:
 [1]
  1. Brookhaven National Lab. (BNL), Upton, NY (United States)
Publication Date:
Research Org.:
Brookhaven National Lab. (BNL), Upton, NY (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Biological and Environmental Research (BER) (SC-23)
OSTI Identifier:
1462437
Report Number(s):
BNL-207916-2018-JAAM
Journal ID: ISSN 1934-3639
Grant/Contract Number:  
SC0012704
Resource Type:
Accepted Manuscript
Journal Name:
Current Protocols in Molecular Biology
Additional Journal Information:
Journal Volume: 124; Journal Issue: 1; Journal ID: ISSN 1934-3639
Publisher:
Wiley
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; BL21(DE3); BL21-AI; MDAG non-inducing growth media; asymmetric ligation; auto-induction; basal expression; co-expression; pAL vectors

Citation Formats

Studier, F. William. T7 Expression Systems for Inducible Production of Proteins from Cloned Genes in E. coli. United States: N. p., 2018. Web. doi:10.1002/cpmb.63.
Studier, F. William. T7 Expression Systems for Inducible Production of Proteins from Cloned Genes in E. coli. United States. doi:10.1002/cpmb.63.
Studier, F. William. Tue . "T7 Expression Systems for Inducible Production of Proteins from Cloned Genes in E. coli". United States. doi:10.1002/cpmb.63. https://www.osti.gov/servlets/purl/1462437.
@article{osti_1462437,
title = {T7 Expression Systems for Inducible Production of Proteins from Cloned Genes in E. coli},
author = {Studier, F. William},
abstractNote = {Inducible T7 expression systems are capable of producing a wide range of proteins in E. coli. Improvements over common practice include: preventing unintended induction by establishing and maintaining expression strains in non-inducing growth media comprised entirely of purified components instead of complex growth media that may variably induce target proteins on approach to saturation; and expressing many target proteins in parallel by convenient and productive auto-induction in BL21(DE3) and other suitable hosts, instead of IPTG induction. From earliest days, basal expression prevented establishment of inducible strains for producing proteins that are stressful to the host. Newly developed pAL vectors now reduce basal expression to levels where coding sequences for even the most stressful proteins can be maintained and induced. In conclusion, asymmetric ligation allows simple and efficient cloning of individual coding sequences or simultaneous cloning of two or three coding sequences for co expression from a single pAL vector.},
doi = {10.1002/cpmb.63},
journal = {Current Protocols in Molecular Biology},
number = 1,
volume = 124,
place = {United States},
year = {2018},
month = {7}
}

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