期刊论文

中文

中南医学科学杂志

2095-1116

2011

39

6

611-614,618

10.15972/j.cnki.43-1509/r.2011.06.004

309701 65、81102230:国家自然科学基金 2008 KS30:衡阳市科技局项目

本校为第一机构

目的 克隆沙眼衣原体(Chlamydia trachomatis,Ct)T细胞抗原Ct143基因,表达并纯化该蛋白,检测其免疫活性. 方法 采用PCR技术从Ct基因组中扩增Ct143基因,并构建pGEX-6p-Ct143原核表达质粒,转化E.coli XL1-Blue,IPTG诱导重组GST-CT143融合蛋白表达,经酶切后得到无GST标签的纯蛋白免疫Balb/c鼠,ELISA及Western-blot分析CT143蛋白免疫原性及免疫反应性. 结果 成功构建了pGEX-6p-CT143原核表达质粒,序列测定证实重组质粒插入片段与GenBank登陆的Ct D型一致,长度为843 bp；GST-CT143融合蛋白在E.coli中获得稳定高效表达；纯蛋白与佐剂混匀后肌肉免疫小鼠,ELISA检测免疫小鼠的血清效价为1∶12 800,Weste...

Objective To clone,express and purify the Chlamydia trachomatis T cell antigen CT143 and preliminarily characterize the antigenicity of purified recombinant protein. Methods The ORF of Ct143 was cloned and inserted into the expression vector pGEX6p-2 to construct pGEX-6p-Ct143 recombinant plasmid. The recombinant plasmid was transformed into E. coli XL1-Blue to express GST-CT143 fusion protein,then purified with Glutathione Sepharose 4B Beads and digested with PreScission Protease to remove the GST tag. The pure protein was used to immunize Bal...