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

Studier, F. William

doi:10.1002/cpmb.63

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

Journal Article · Tue Jul 17 00:00:00 EDT 2018 · Current Protocols in Molecular Biology

DOI:https://doi.org/10.1002/cpmb.63· OSTI ID:1462437

Studier, F. William ^[1]

Brookhaven National Lab. (BNL), Upton, NY (United States)

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.

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Research Organization:: Brookhaven National Laboratory (BNL), Upton, NY (United States)

Sponsoring Organization:: USDOE Office of Science (SC), Biological and Environmental Research (BER)

Grant/Contract Number:: SC0012704

OSTI ID:: 1462437

Report Number(s):: BNL-207916-2018-JAAM

Journal Information:: Current Protocols in Molecular Biology, Vol. 124, Issue 1; ISSN 1934-3639

Publisher:: WileyCopyright Statement

Country of Publication:: United States

Language:: English

References (23)

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Construction of a strain-producer of the chimeric protein consisting of RNA polymerase and a DNA-affinity domain Kazlovskiy, I. S.; Zinchenko, M. A. Doklady of the National Academy of Sciences of Belarus, Vol. 62, Issue 5 https://doi.org/10.29235/1561-8323-2018-62-5-601-607	journal	October 2018

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59 BASIC BIOLOGICAL SCIENCES
BL21(DE3)
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MDAG non-inducing growth media
asymmetric ligation
auto-induction
basal expression
co-expression
pAL vectors

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

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References (23)

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