Human Era protein in Escherichia coli Expression and Purification.docVIP

 PAGE \* MERGEFORMAT 14 Human Era protein in Escherichia coli Expression and Purification Author: Wu Yuan-Ming, CHEN Su-min, CHEN Nan-Chun, Zong-Ling, Hui-Ping Liu, Junjie [Keywords:] human genome Era Keywords:: Human Era gene; gene expression; protein purification Abstract: The purpose of Escherichia coli in the era of human gene (referred to as hera) for expression and purification. Hera gene amplification method of PCR, sequencing, and cloned into the E. coli fusion expression vector pRSET-C, recombinant plasmid to E. coli pRSETC-hera BL21 (DE3) as host bacteria with IPTG induced for expression. then use the method of affinity chromatography purified protein expression. The results of the hera gene cloned and sequenced correctly; the use of vector pRSET-C fusion expression in E. coli the entire long-hera-cDNA gene expression accounted for 59.6% of total bacterial proteins. fusion protein in bacteria in vivo mainly in the form of inclusion bodies in denaturing conditions using Ni-NTA affinity column purification of the fusion protein, its purity reached 98.0 %. Conclusion The use of genetic recombination technology in E. coli highly expressed genes of the hera. and carried out a preliminary purification of fusion protein. Keywords: human Era gene; gene expression; protein purifi-cation Abstract: AIM To express human Era protein in E.coli and purify it preliminarily.METHODS After being amplified by PCR and identified by sequencing the human era gene was in-serted into expression vector pRSET-C.The recombinant plasmid pRSETC-hera was transformed into BL21 (DE3) and induced with IPTG.The expressed protein was purified by affinity chromotography.RESULTS The human era gene was amplified and the sequence proved to be correct.The hera-cDNA gene had been cloned into the vector and ex-pressed in E.coli, and the expressed protein took59.6% of the total bacterial protein.The fused protein existed in the pattern of inclusion body, and it was purified on denatur