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Rapid evolution of regulatory element libraries for tunable transcriptional and translational control of gene expression

查看全文 作　　者：Erqing [1,2]Jin;Lynn [1]Wong;Yun [1]Jiao;Jake [1]Engel;Benjamin [1]Holdridge;Peng [1]Xu 高影响力作者 机构地区：[1]Department of Chemical,Biochemical and Environmental Engineering,University of Maryland Baltimore County,1000 Hilltop Circle,Baltimore,MD 21250,United States;[2]Department of Food Science and Engineering,Jinan University,601 West Huangpu Road,Guangzhou 510632,China高影响力机构 出　　处：《Synthetic and Systems Biotechnology》索引2017年第2卷第4期,共7页高影响力期刊 基　　金：The authors would like to acknowledge the Department of Chemical,Biochemical and Environmental Engineering,College of Engineering and Information Technology,Office of the Vice President for Research(gratn number 10145-1113-021-STRT7XUPMAIN)at University of Maryland Baltimore County for funding support. 摘　　要：Engineering cell factories for producing biofuels and pharmaceuticals has spurred great interests to develop rapid and efficient synthetic biology tools customized for modular pathway engineering.Along the way,combinatorial gene expression control through modification of regulatory element offered tremendous opportunity for fine-tuning gene expression and generating digital-like genetic circuits.In this report,we present an efficient evolutionary approach to build a range of regulatory control elements.The reported method allows for rapid construction of promoter,5'UTR,terminator and trans-activating RNA libraries.Synthetic overlapping oligos with high portion of degenerate nucleotides flanking the regulatory element could be efficiently assembled to a vector expressing fluorescence reporter.This approach combines high mutation rate of the synthetic DNA with the high assembly efficiency of Gibson Mix.Our constructed library demonstrates broad range of transcriptional or translational gene expression dynamics.Specifically,both the promoter library and 50UTR library exhibits gene expression dynamics spanning across three order of magnitude.The terminator library and trans-activating RNA library displays relatively narrowed gene expression pattern.The reported study provides a versatile toolbox for rapidly constructing a large family of prokaryotic regulatory elements.These libraries also facilitate the implementation of combinatorial pathway engineering principles and the engineering of more efficient microbial cell factory for various biomanufacturing applications. 关 键 词：PROMOTER 50UTR TERMINATOR Riboregulatory Trans-activating RNA Library Synthetic biology
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