title: Rapid Diversification of BetI-Based Transcriptional Switches for the Control of Biosynthetic Pathways and Genetic Circuits.
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.
authors.affiliation: Department of Applied Chemistry and Biotechnology, Faculty of Engineering, Chiba University , 1-33 Yayoi-Cyo, Inage-ku, Chiba 263-8522, Japan.
authors.forename: Kazuya
authors.initials: K
authors.lastname: Saeki
authors.affiliation: Department of Applied Chemistry and Biotechnology, Faculty of Engineering, Chiba University , 1-33 Yayoi-Cyo, Inage-ku, Chiba 263-8522, Japan.
authors.forename: Masahiro
authors.initials: M
authors.lastname: Tominaga
authors.affiliation: Department of Applied Chemistry and Biotechnology, Faculty of Engineering, Chiba University , 1-33 Yayoi-Cyo, Inage-ku, Chiba 263-8522, Japan.
authors.forename: Shigeko
authors.initials: S
authors.lastname: Kawai-Noma
authors.affiliation: Department of Applied Chemistry and Biotechnology, Faculty of Engineering, Chiba University , 1-33 Yayoi-Cyo, Inage-ku, Chiba 263-8522, Japan.
authors.forename: Kyoichi
authors.initials: K
authors.lastname: Saito
authors.affiliation: Department of Applied Chemistry and Biotechnology, Faculty of Engineering, Chiba University , 1-33 Yayoi-Cyo, Inage-ku, Chiba 263-8522, Japan.|Precursory Research for Embryonic Science and Technology (PRESTO), Japan Science and Technology Agency (JST), 4-1-8 Honcho, Kawaguchi, Saitama 332-0012, Japan.
authors.forename: Daisuke
authors.initials: D
authors.lastname: Umeno
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D053898:Biosynthetic Pathways; D002794:Choline / Q000737:chemistry; D004926:Escherichia coli / Q000235:genetics; D053263:Gene Regulatory Networks; D011401:Promoter Regions, Genetic; D058615:Synthetic Biology / Q000379:methods*; D014157:Transcription Factors / Q000235:genetics* / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D014157:Transcription Factors; D002794:Choline
keywords: Boolean logic gates; carotenoid; directed evolution; genetic switch; induction system; liquid-handling; lycopene; operon; selection