首页|期刊导航|高技术通讯（英文版）|Overexpression and export of Vibrio anguillarum metalloprotease in Escherichia coli
高技术通讯（英文版）2007，Vol.13Issue(1)：79-84,6.
Overexpression and export of Vibrio anguillarum metalloprotease in Escherichia coli
Overexpression and export of Vibrio anguillarum metalloprotease in Escherichia coli
摘要
Abstract
Vibrio anguillarum metalloprotease, an extracellular zinc metalloprotease involved in the virulence mechanism of Vibrio anguillarum, is synthesized from the empA gene as a 611-residue precursor and naturally secreted via Sec secretion pathway in Vibrio anguillarum. In this study, heterologous expression of the empA gene encoding metallopmtease and export of the recombinant metalloprotease in Escherichia coliwere examined. The empA gene was subcloned into pBAD24 with arabinose promoter and sequenced. The sequence encoded a polypeptide(611 amino acids)consisting of four domains: a signal peptide, an Nterminal propeptide, a mature region and a C-terminal propeptide. The empA gene inserted in plasmid pBAD24 was overexpressed in TOP10 strain of E. Coli after arabinose induction. The 36kDa polypeptide of the recombinant metalloprotease as the mature protease was further confirmed by SDS-PAGE and immunoblotting. It was found that recombinant metalloprotease with the EmpA activity and antigenicity wasexported into the periplasm of Escherichia coli cells via Sec translocation pathway, whereas it was secreted into extracellular environments in V. Anguillarum. The results imply that the expression, export and processing mechanism of the protein in E. Coli are similar to those in V. Anguillarum.关键词
Vibrio anguillarum/metalloprotease/overexpression/export/translocation/Escherichia coli/periplasmKey words
Vibrio anguillarum/metalloprotease/overexpression/export/translocation/Escherichia coli/periplasm分类
医药卫生引用本文复制引用
Zhang Fengli,Chi Zhenming,Chen Jixiang,Wu Longfei,Liang Likun..Overexpression and export of Vibrio anguillarum metalloprotease in Escherichia coli[J].高技术通讯（英文版）,2007,13(1):79-84,6.基金项目
Supported by the National Natural Science Foundation of China(No.30328021). （No.30328021）
