title: Construction of a host-independent T7 expression system with small RNA regulation.
abstract: It is desirable to build a universal and efficient protein expression system for wild-type prokaryotic strains in biotechnology industry and the outstanding T7 expression system could be a good candidate. However, the current utilization of T7 system depends on the specific DE3 lysogenic hosts, which severely limits its application in wild-type strains. In this study, a host-independent T7 expression system without relying on DE3 lysogenic hosts to provide T7 RNA Polymerase was developed. T7 RNA Polymerase gene (Gene1) and T7 Promoter were successfully integrated into a single plasmid with the regulation of proper antisense RNA to limit T7 RNA Polymerase expression at a non-lethal level. This host-independent T7 expression system realized efficient protein expression in 4 non-DE3 Escherichia coli strains and a wild-type Sinorhizobium strain TH572.
authors.affiliation: Key Laboratory for Industrial Biocatalysis, Ministry of Education, Department of Chemical Engineering, Tsinghua University, Beijing 100084, China.
authors.forename: Gang
authors.initials: G
authors.lastname: Wang
authors.affiliation: Key Laboratory for Industrial Biocatalysis, Ministry of Education, Department of Chemical Engineering, Tsinghua University, Beijing 100084, China. Electronic address: liqiang@tsinghua.edu.cn.
authors.forename: Qiang
authors.initials: Q
authors.lastname: Li
authors.affiliation: Key Laboratory for Industrial Biocatalysis, Ministry of Education, Department of Chemical Engineering, Tsinghua University, Beijing 100084, China.
authors.forename: Dikai
authors.initials: D
authors.lastname: Xu
authors.affiliation: Key Laboratory for Industrial Biocatalysis, Ministry of Education, Department of Chemical Engineering, Tsinghua University, Beijing 100084, China.
authors.forename: Mingxin
authors.initials: M
authors.lastname: Cui
authors.affiliation: Key Laboratory for Industrial Biocatalysis, Ministry of Education, Department of Chemical Engineering, Tsinghua University, Beijing 100084, China.
authors.forename: Xiao
authors.initials: X
authors.lastname: Sun
authors.affiliation: Key Laboratory for Industrial Biocatalysis, Ministry of Education, Department of Chemical Engineering, Tsinghua University, Beijing 100084, China.
authors.forename: Yanyan
authors.initials: Y
authors.lastname: Xu
authors.affiliation: College of Life Science and Technology, Beijing University of Chemical Technology, Beijing 100029, China.
authors.forename: Wenya
authors.initials: W
authors.lastname: Wang
mesh_terms: D017123:Bacteriophage T7 / Q000235:genetics*; D012321:DNA-Directed RNA Polymerases / Q000235:genetics*; D016372:RNA, Antisense / Q000235:genetics; D014764:Viral Proteins / Q000235:genetics*
chemical_list: D016372:RNA, Antisense; D014764:Viral Proteins; C062210:bacteriophage T7 RNA polymerase; D012321:DNA-Directed RNA Polymerases
keywords: Antisense RNA; Host-independent expression system; T7 Promoter; T7 RNA Polymerase