To investigate the roles of HtrA protein in bacterial adaption to stresses, htra gene was cloned from an Escherichia coli strain isolated from silver fox, double digested, and ligated into the prokaryotic expression v...To investigate the roles of HtrA protein in bacterial adaption to stresses, htra gene was cloned from an Escherichia coli strain isolated from silver fox, double digested, and ligated into the prokaryotic expression vector pET32a. Then, the recombinant plasmid pET32a-htra was transformed into E. coli BL21 competent cells. The over-expression of His-HtrA protein was induced by the addition of IPTG at a final concentration of 1 mmol/L, and verified by SDA-PAGE and Western blot. In addition, the protein was purified with Ni-NTA agarose, and used to immunize mice,and the polyclonal antibody specifically bound to E. coli HtrA protein. The results will provide a theoretical basis for studying the roles of HtrA protein and developing vaccines for fur-bearing animals against E. coli.展开更多
基金Supported by China Spark Program(2015GA620002)Key Technologies Research and Development Program of Hebei Province(14826613D)+2 种基金PhD Start-up Fund of Hebei Normal University of Science and Technology(2015YB002)Scientific and Technological Research Program of Institutions of Higher Education of Hebei Province(ZD2016067)the Fund of Qinhuangdao Academy of Agricultural Sciences(2014-04)
文摘To investigate the roles of HtrA protein in bacterial adaption to stresses, htra gene was cloned from an Escherichia coli strain isolated from silver fox, double digested, and ligated into the prokaryotic expression vector pET32a. Then, the recombinant plasmid pET32a-htra was transformed into E. coli BL21 competent cells. The over-expression of His-HtrA protein was induced by the addition of IPTG at a final concentration of 1 mmol/L, and verified by SDA-PAGE and Western blot. In addition, the protein was purified with Ni-NTA agarose, and used to immunize mice,and the polyclonal antibody specifically bound to E. coli HtrA protein. The results will provide a theoretical basis for studying the roles of HtrA protein and developing vaccines for fur-bearing animals against E. coli.
