Engineering of a Promoter Repressed by a Light-Regulated Transcription Factor in Escherichia coli

作     者：Daniel Camsund Alfonso Jaramillo Peter Lindblad 

作者机构：Microbial ChemistryDepartment of Chemistry-ÅngströmUppsala UniversityUppsalaSweden Molecular Systems BiologyDepartment of Cell and Molecular BiologyUppsala UniversityUppsalaSweden Warwick Integrative Synthetic Biology Centre(WISB)and School of Life SciencesUniversity of WarwickCoventryUK Génomique MétaboliqueGenoscopeInstitut François JacobCEACNRSUniv EvryUniversitéParis-SaclayEvryFrance Institute for Integrative Systems Biology(I2SysBio)CSIC–Universitat de València46980 PaternaSpain 

基　　金：the fluorescence spectrum of EL222,and Adam Wegelius is gratefully acknowledged for experimental assistance.AJ acknowledges funding from FP7-ICT-610730(EVOPROG) FP7-KBBE-613745(PROMYS) H2020 Marie Sklodowska-Curie Actions 642738(MetaRNA) Biotechnology and Biological Biological Sciences Research Council(BBSRC)BB/P020615/1(EVO-ENGINE),Engineering and Physical Sciences Research Council-Biotechnology and Biological Sciences Research Council(EPSRC-BBSRC)BB/M017982/1(WISB centre),and the departmental allocation from the School of Life Sciences(U.Warwick) PL acknowledges funding from the Knut och Alice Wallenbergs Stiftelse(project MoSE,No.2011.0067)and the Swedish Energy Agency(No.11674-5) 

出 版 物：《BioDesign Research》 (生物设计研究（英文）)

年 卷 期：2021年第2021卷第1期

页      码：59-69页

摘      要：Light-regulated gene expression systems allow controlling gene expression in space and time with high *** to previous synthetic light sensors that incorporate two-component systems which require localization at the plasma membrane,soluble one-component repression systems provide several advantageous ***,they are soluble and able to diffuse across the ***,they are smaller and of lower complexity,enabling less taxing expression and optimization of fewer ***,repression through steric hindrance is a widespread regulation mechanism that does not require specific interaction with host factors,potentially enabling implementation in different ***,we present the design of the synthetic promoter P_(EL)that in combination with the light-regulated dimer EL222 constitutes a onecomponent repression *** by previously engineered synthetic promoters and the Escherichia coli lacZYA promoter,we designed P_(EL)with two EL222 operators positioned to hinder RNA polymerase binding when EL222 is bound.P_(EL)is repressed by EL222 under conditions of white light with a light-regulated repression ratio of ***,alternating conditions of darkness and light in cycles as short as one hour showed that repression is *** design of the P_(EL)EL222 system herein presented could aid the design and implementation of analogous one-component optogenetic repression ***,we compare the P_(EL)-EL222 system with similar systems and suggest general improvements that could optimize and extend the functionality of EL222-based as well as other one-component repression systems.

主 题 词：pressed system analogous 

学科分类：07[理学] 0701[理学-数学类] 070101[070101] 

D　O　I：10.34133/2021/9857418

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