猪链球菌2型荚膜多糖cps2J基因的克隆与原核表达  

Expression of cps2J fusion gene of Streptococcus suis serotype 2 in E.coli

作　　者：刘燕[1] 刘军[1] 冯书章[1] 郭学军[1] 孙洋[1] 祝令伟[1] 

机构地区：[1]军事医学科学院军事兽医研究所,吉林长春130062

出　　处：《中国预防兽医学报》2009年第6期434-437,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家"973"计划项目(2006CB504402);吉林省科技发展计划资助项目(20080130)

摘　　要：利用PCR方法从猪链球菌2型(SS2)四川资阳分离株449-1扩增出cps2J基因,克隆到pMD18-T载体,经酶切与表达质粒pMAL-p2x构建重组表达质粒pMALp2x-cps2J,转化至宿主菌TB1中诱导表达。SDS-PAGE电泳检测结果表明,重组菌株表达出了约80ku的可溶性目的蛋白MBP-cps2J,约占菌体蛋白总量的13.5%。表达蛋白用Amylose树脂层析柱纯化,将纯化的融合蛋白MBP-cps2J免疫家兔制备抗血清,经ELISA和协同凝集试验检测,其与SS2呈特异性阳性反应。To study the immunogenicity ofcps2J gene of Streptococcus suis serotype 2 （SS2）, the cps2J gene was PCR- amplified from SS2 449-1 strain from Sichuan, and cloned into expression vector pMAL-p2x. The recombinant plasmid was transformed into E.coli TB1 and induced by IPTG for protein expression. The fusion protein MBP-cps2J was expressed in a soluble form, with an approximate yield 13.5 % of the whole bacteria protein. The fusion protein was purified and used to immunize the rabbits. High antibody titer （1：16 000） was detected by ELISA. The antisera collected from the immunized rabbits reacted specifically with SS2.

关 键 词：猪链球菌2型 cps2J基因 协同凝集试验 

分 类 号：S852.611[农业科学—基础兽医学]