期刊论文

中文

免疫学杂志

1000-8861

2011

27

2

93-97,104

10.13431/j.cnki.immunol.j.20110022

国家自然科学基金（30970165） 湖南省自然科学基金重点项目（09JJ3059）

本校为第一机构

目的分析沙眼衣原体隐蔽性质粒编码的质粒蛋白pORF5在感染细胞中的定位并初步探讨免疫原性特征。方法 PCR扩增pORF5质粒蛋白编码基因,构建原核表达重组体pGEX-6p/pORF5,重组体转化大肠杆菌XL1-blue中诱导表达融合蛋白;融合蛋白经Glutathione Sepharose亲和层析纯化后免疫小鼠,制备单克隆抗体和多克隆抗体,间接免疫荧光技术及免疫印迹鉴定抗体的特异性,并分析pORF5质粒蛋白在感染细胞中的分布特征。采用ELISA方法分析pORF5质粒蛋白的免疫原性。结果 pORF5原核表达重组体成功构建,融合蛋白在大肠杆菌中可溶性表达;pORF5主要分布于宿主细胞胞浆,但也少量分布在EB、RB上;pORF5能与衣原体患者血清及鼠...

We aimed to localize the plasmid protein pORF5 in chlamydia infected cells and investigate the immunogenicity of pORF5. Firstly, the open reading frame encoding for pORF5 protein from the Chlamydia trachomatis plasmid was amplified and cloned into the pGEX-6p vector. Then the recombinant plasmid pGEX-6p/pORF5 was transformed into XL1-blue E.coli with glutathione-s-transferase(GST) to express fusion protein. After purified with Glutathione Sepharose 4B beads, the pORF5 fusion protein was used to immunize mice to make monoclonal antibody and polyclonal antibody, which were subsequently used to l...