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Title 

Tunable control of an Escherichia coli expression system for the overproduction of membrane proteins by titrated expression of a mutant lac repressor

 

LacI 변이체의 정밀 조절을 통한 대장균 내 막단백질의 과량 생산

Authors 

Seong Keun KimDae-Hee LeeOh Cheol KimJ F KimS H Yoon

Publisher 

American Chemical Society

Issue Date 

2017

Citation 

ACS Synthetic Biology

Keywords 

Escherichia coliexpression systemlac repressormembrane proteinmutant LacI

Abstract 

Most inducible expression systems suffer from growth defects, leaky basal induction, and inhomogeneous expression levels within a host cell population. These difficulties are most prominent with the overproduction of membrane proteins that are toxic to host cells. Here, we developed an Escherichia coli inducible expression system for membrane protein production based on titrated expression of a mutant lac repressor (mLacI). Performance of the mLacI inducible system was evaluated in conjunction with commonly used lac operator-based expression vectors using a T7 or tac promoter. Remarkably, expression of a target gene can be titrated by the dose-dependent addition of l-rhamnose, and the expression levels were homogeneous in the cell population. The developed system was successfully applied to overexpress three membrane proteins that were otherwise difficult to produce in E. coli. This gene expression control system can be easily applied to a broad range of existing protein expression systems and should be useful in constructing genetic circuits that require precise output signals

ISSN 

2161-5063

Link 

http://dx.doi.org/10.1021/acssynbio.7b00102

Appears in Collections

1. Journal Articles > Journal Articles

Registered Date

2019-05-02


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