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Construction of a host-independent T7 expression system with small RNA regulation

Wang, Gang, Li, Qiang, Xu, Dikai, Cui, Mingxin, Sun, Xiao, Xu, Yanyan, Wang, Wenya
Journal of biotechnology 2014 v.189 pp. 72-75
DNA-directed RNA polymerase, Escherichia coli, Sinorhizobium, antisense RNA, biotechnology, genes, hosts, industry, plasmids, protein synthesis
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.