大肠杆菌RNaseⅢ基因的克隆与表达

Cloning and Expression of Escherichia coli RNaseⅢ

目的:克隆并在原核表达系统中表达RNaseⅢ基因。方法:提取大肠杆菌JM109株的基因组DNA,以之为模板扩增得到RNaseⅢ基因全序列,将该编码序列克隆到原核表达载体pET-22b(+)中,转化大肠杆菌BL21(DE3),IPTG诱导重组RNaseⅢ表达。结果与结论:在大肠杆菌中克隆到了RNaseⅢ的全基因,经测序证明与数据库中报道的序列一致;表达的重组RNaseⅢ主要以包涵体形式存在。

Objective：To clone and prokaryotic express the RNaseⅢ gene of Escherichia coli.Methods：The genomic DNA was extracted from E.coli strain JM109,and RNaseⅢ gene was amplified based on it,and was then inserted into a prokaryotic expression vector named pET-22b（＋）.The recombinant plasmids were transformed into E.coli BL21（DE3） and induced with IPTG.Results and Conclusion：The amplified RNaseⅢ gene was confirmed by DNA sequence analysis.The RNaseⅢ protein was successfully expressed in form of inclusion body.