Visualized and precise design of artificial small RNAs for regulating T7 RNA polymerase and enhancing recombinant protein folding in Escherichia coli  

作　　者：Yujia Zhao Jingjing Fan Jinlin Li Jun Li Xiaohong Zhou Chun Li 

机构地区：[1]Department of Biological Engineering,School of Life Science,Beijing Institute of Technology,Beijing 10008,China

出　　处：《Synthetic and Systems Biotechnology》2016年第4期265-270,共6页合成和系统生物技术（英文）

基　　金：The author would like to acknowledge the National Science Fund for Distinguished Young Scholars(21425624);the National Natural Science Foundation of China(21476026,21376028).

摘　　要：Small non-coding RNAs(sRNAs)have received much attention in recent years due to their unique biological properties,which can efficiently and specifically tune target gene expressions in bacteria.Inspired by natural sRNAs,recent works have proposed the use of artificial sRNAs(asRNAs)as genetic tools to regulate desired gene that has been applied in several fields,such as metabolic engineering and bacterial physiology studies.However,the rational design of asRNAs is still a challenge.In this study,we proposed structure and length as two criteria to implement rational visualized and precise design of asRNAs.T7 expression system was one of the most useful recombinant protein expression systems.However,it was deeply limited by the formation of inclusion body.To settle this problem,we designed a series of asRNAs to inhibit the T7 RNA polymerase(Gene1)expression to balance the rate between transcription and folding of recombinant protein.Based on the heterologous expression of Aspergillus oryzae Li-3 glucuronidase in E.coli,the asRNA-antigene1-17bp can effectively decrease the inclusion body and increase the enzyme activity by 169.9%.

关 键 词：Artificial small RNAs Visualized and precise design Prokaryotic T7 expression system Inclusion body 

分 类 号：R73[医药卫生—肿瘤]