Rapid Diversification of BetI-Based Transcriptional Switches for the Control of Biosynthetic Pathways and Genetic Circuits

Kazuya Saeki, Masahiro Tominaga, Shigeko Kawai-Noma, Kyoichi Saito, Daisuke Umeno

Research output: Contribution to journalArticlepeer-review

8 Citations (Scopus)

Abstract

Synthetic biologists are in need of genetic switches, or inducible sensor/promoter systems, that can be reliably integrated in multiple contexts. Using a liquid-based selection method, we systematically engineered the choline-inducible transcription factor BetI, yielding various choline-inducible and choline-repressive promoter systems with various input-output characteristics. In addition to having high stringency and a high maximum induction level, they underwent a graded and single-peaked response to choline. Taking advantage of these features, we demonstrated the utility of these systems for controlling the carotenoid biosynthetic pathway and for constructing two-input logic gates. Additionally, we demonstrated the rapidity, throughput, robustness, and cost-effectiveness of our selection method, which facilitates the conversion of natural genetic controlling systems into systems that are designed for various synthetic biology applications.

Original languageEnglish
Pages (from-to)1201-1210
Number of pages10
JournalACS Synthetic Biology
Volume5
Issue number11
DOIs
Publication statusPublished - 2016 Nov 18
Externally publishedYes

Keywords

  • Boolean logic gates
  • carotenoid
  • directed evolution
  • genetic switch
  • induction system
  • liquid-handling
  • lycopene
  • operon
  • selection

ASJC Scopus subject areas

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

Fingerprint Dive into the research topics of 'Rapid Diversification of BetI-Based Transcriptional Switches for the Control of Biosynthetic Pathways and Genetic Circuits'. Together they form a unique fingerprint.

Cite this