Evolutionary Design of Choline-Inducible and -Repressible T7-Based Induction Systems

Kohei Ike, Yusuke Arasawa, Satoshi Koizumi, Satoshi Mihashi, Shigeko Kawai-Noma, Kyoichi Saito, Daisuke Umeno*

*この研究の対応する著者

研究成果: Article査読

12 被引用数 (Scopus)

抄録

By assembly and evolutionary engineering of T7-phage-based transcriptional switches made from endogenous components of the bet operon on the Escherichia coli chromosome, genetic switches inducible by choline, a safe and inexpensive compound, were constructed. The functional plasticity of the BetI repressor was revealed by rapid and high-frequency identification of functional variants with various properties, including those with high stringency, high maximum expression level, and reversed phenotypes, from a pool of BetI mutants. The plasmid expression of BetI mutants resulted in the choline-inducible (Bet-ON) or choline-repressible (Bet-OFF) switching of genes under the pT7/betO sequence at unprecedentedly high levels, while keeping the minimal leaky expression in uninduced conditions.

本文言語English
ページ(範囲)1352-1360
ページ数9
ジャーナルACS Synthetic Biology
4
12
DOI
出版ステータスPublished - 2015 12月 18
外部発表はい

ASJC Scopus subject areas

  • 生体医工学
  • 生化学、遺伝学、分子生物学(その他)

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