中国农业科学院农业信息研究所 中国农学会
Agricultural Information Institute of CAAS China Association of Agricultural Science Societies
En
Ch
中国农业科学院农业信息研究所 中国农学会 主办

AI检索

订阅

返回顶部
当前位置: 首页 > 文章 > 福氏志贺菌2型外膜蛋白A的原核表达及多克隆抗体的制备 动物医学进展 2015,36 (5) 6-10
Position: Home > Articles > Prokaryotic Expression and Polyclonal Antibody Preparation of Shigella flexneri 2a Outer Membrane Protein A Progress in Veterinary Medicine 2015,36 (5) 6-10

福氏志贺菌2型外膜蛋白A的原核表达及多克隆抗体的制备

作  者：
刘祥
单  位：
陕西理工学院生物科学与工程学院
关键词：
福氏志贺菌;OmpA蛋白;多克隆抗体;酶联免疫吸附试验;Western blot
摘  要：
福氏志贺菌外膜蛋白A是细菌入侵宿主细胞的作用蛋白,同时激活机体的免疫机制对抗细菌的感染,在动物疫苗上有很好的应用前景。采用分子克隆方法获得福氏志贺菌OmpA蛋白表达菌株;利用SDS-PAGE电泳切胶纯化、尿素梯度复性获得OmpA蛋白,免疫小鼠制备OmpA蛋白多克隆抗体;采用ELISA法检测抗体滴度,Western blot检测抗血清特异性;DNAMan与MEGA软件分析OmpA蛋白系统进化关系。结果显示,OmpA重组载体双酶切、DNA测序鉴定结果,以及蛋白表达、纯化条带大小与预测一致;ELISA法确认OmpA抗血清滴度达1∶1 600,Western blot证实抗血清具有很好的特异性。OmpA序列系统发生分析发现不同种的致病菌存在同源性,尤其C-端同源性较高,并且不同种志贺菌具有更高的同源性。表明成功制备OmpA蛋白和小鼠多克隆抗体,为OmpA蛋白功能与疫苗开发研究奠定了基础。
译  名：
Prokaryotic Expression and Polyclonal Antibody Preparation of Shigella flexneri 2a Outer Membrane Protein A
作  者：
LIU Xiang;College of Biological Sciences and Engineering,Shaanxi University of Technology;
关键词：
Shigella flexneri;;ompA protein;;polyclonal antibody;;ELISA;;Western blot
摘  要：
Shigella flexneri OmpA(outer membrane protein A)serves as a receptor for bacteria to invade host cells,while it opposes bacterium infection by inducing immunologic mechanism,and has important perspective in animal vaccine.The S.flexneri OmpA protein expression strain was obtained by molecular clone;OmpA was purified by the way of SDS-PAGE gel slices,renatured by urea gradient,and used to immunize mice to prepare the polyclonal antibodies;The antibody titer and specificity were detected by ELISA and Western blot,respectively.DNA Man and MEGA softwares were used to analyze phylogenetic relationships of OmpA.The results showed that OmpA recombinant vector was digested and sequenced to confirm the correct construction,and OmpA expression and purification of strip size agreed with the prediction.The OmpA antibody titer was 1∶1 600 by ELISA,and Western blot proved that the antiserum had good specificity.OmpA Phylogenetic analysis found that the different kinds of pathogenic bacteria had homologous,especially C-end homology was high,and different kinds of Shigellaspecies had higher homology.Then,Succession of cloning of OmpA expression vector,OmpA purification,preparation and identification of the polyclonal antibodies of OmpA,and analyses of genetic evolution relationship of OmpA sequence were done.The work laid the foundation for the protein function and vaccine development of OmpA.

相似文章

导出引用
计量
文章访问数: 13
HTML全文浏览量: 0
PDF下载量: 0

所属期刊

推荐期刊

友情链接Friendly link
农业信息研究所Agricultural information institute 农业智能知识服务平台Agricultural intelligent knowledge service platform 农业学术服务平台Agricultural Knowledge Service System 中国知网CNKI 维普网VIP 万方WANFANG DATA
联系我们Contact us

电话Tell：010-82109652-808

邮箱E-mail：agrijournal@caas.cn

服务Service
关于我们About us 意见建议Opinions and suggest
版权说明Copyright
单位：中国农业科学院农业信息研究所 中国农学会 © 2017中国农业科学院农业信息研究所 ICP备案号：京ICP备09089781号-46