Tunable Control of an Escherichia coli Expression System for the Overproduction of Membrane Proteins by Titrated Expression of a Mutant lac Repressor

Seong Keun Kim, Dae Hee Lee, Oh Cheol Kim, Jihyun F. Kim, Sung Ho Yoon

Research output: Contribution to journalArticle

4 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)1766-1773
Number of pages8
JournalACS Synthetic Biology
Volume6
Issue number9
DOIs
Publication statusPublished - 2017 Sep 15

Fingerprint

Lac Repressors
Escherichia coli
Membrane Proteins
Proteins
Membranes
Cells
Rhamnose
Poisons
Gene expression
Population
Genes
Control systems
Gene Expression
Defects
Networks (circuits)
Growth

All Science Journal Classification (ASJC) codes

  • Biomedical Engineering
  • Biochemistry, Genetics and Molecular Biology (miscellaneous)

Cite this

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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.",
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Tunable Control of an Escherichia coli Expression System for the Overproduction of Membrane Proteins by Titrated Expression of a Mutant lac Repressor. / Kim, Seong Keun; Lee, Dae Hee; Kim, Oh Cheol; Kim, Jihyun F.; Yoon, Sung Ho.

In: ACS Synthetic Biology, Vol. 6, No. 9, 15.09.2017, p. 1766-1773.

Research output: Contribution to journalArticle

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AU - Yoon, Sung Ho

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