肺炎链球菌毒力因子PspA的原核表达、纯化及鉴定  

Expression and purification of streptococcus pneumoniae virulence factor PspA

作　　者：胥文春[1] 曹炬[1] 许颂霄[1] 罗进勇[1] 阳强[1] 尹一兵[1] 

机构地区：[1]重庆医科大学医学检验系实验诊断教研室临床检验诊断学省部共建教育部重点实验室,重庆400016

出　　处：《重庆医科大学学报》2007年第6期571-574,共4页Journal of Chongqing Medical University

基　　金：国家自然科学基金资助项目(30471873)

摘　　要：目的:通过构建原核表达载体,获得肺炎链球菌毒力因子PspA重组蛋白。方法:分离培养肺炎链球菌TIGR4,获取其染色体DNA,采用基因体外重组法将PspA序列克隆到原核表达载体PET-32(a)上,用酶切及测序鉴定。将重组质粒转化到大肠杆菌BL21(DE3)中,经IPTG诱导,进行蛋白纯化,获得的重组蛋白用SDS-PAGE和Western blot鉴定。结果:克隆的DNA序列与GenBank中的数据相符,获得的重组蛋白PspA纯度达到90%。结论:成功表达和纯化了PspA,为肺炎链球菌多肽联合疫苗研制和该蛋白的结构生物学分析奠定了基础。Objective：To obtain purified PspA produced by prokaryotic expression system. Methods：Template DNA was isolated from cultured Streptococcus pneumonia TIGR4.By gene recombination technology in vitro,sequence encoding PspA antigen epitope was cloned into PET-32 （a） expression vector. After being confirmed by sequencing,expressed antigen protein was identified by SDS-PAGE and Western blot. Results：The DNA sequence analysis confirmed that the cloned PspA gene was according to GenBank data.The PspA fusion protein was proved by Western blot. Conclusion：A highly expressed recombinant PspA protein was successfully obtained.

关 键 词：肺炎链球菌 PSPA 疫苗 

分 类 号：R392.1[医药卫生—免疫学]