重组大肠埃希菌外膜蛋白C的表达、纯化及多克隆抗体制备  

Expression,purification and polyclonal antibody preparation of recombinant E. coli outer membrane protein OmpC

作　　者：俱雄 陈春琳[1] 刘祥[1] 王成祥 王令[1] 吴三桥[1] 张涛[1] 

机构地区：[1]陕西理工学院生物科学与工程学院,陕西汉中723000 [2]安徽省肥西县特殊教育学校,安徽合肥231200

出　　处：《陕西理工学院学报（自然科学版）》2015年第3期52-57,共6页Journal of Shananxi University of Technology:Natural Science Edition

基　　金：陕西省教育厅科学研究计划项目(2013JK0723);陕西理工学院博士科研启动项目(SLGQD13-15)

摘　　要：采用分子克隆方法获得大肠埃希菌外膜蛋白Omp C表达菌株;利用SDS-PAGE电泳切胶纯化、尿素梯度复性获得Omp C蛋白,免疫小鼠制备Omp C蛋白多克隆抗体。ELISA法证实Omp C抗血清滴度达1∶6 400倍,Western blotting发现Omp C抗血清具有很好的特异性。通过DNAMAN软件对Omp C序列同源性分析显示不同细菌间存在较高同源性,采用MEGA软件对Omp C的系统进化分析发现肠道致病菌亲缘关系更近。为Omp C蛋白免疫学功能研究奠定基础。E.coli outer membrane protein （ OmpC） could oppose bacterium infection by inducing body immunologic mechanism, and had important perspective in vaccine development.The OmpC protein expression strain was obtained by molecular clone;OmpC was purified by the way of SDS-PAGE gel extraction and urea gradient renaturation, and was used to immunize mice to prepare the polyclonal antibody.The OmpC antibody titer was 1∶6 400 by ELISA, and Western blotting proved that the antiserum had good specificity. OmpC homology analysis by DNAMAN showed that different bacteria had high homology;Phylogenetic analysis of OmpC by MEGA proved the close relationship between intestinal tract pathogenic bacteria.These studies laid the groundwork for the immunological and functional research of OmpC.

关 键 词：OmpC蛋白 酶联试验 多克隆抗体 WESTERN BLOTTING 

分 类 号：Q789[生物学—分子生物学]