期刊论文

Wu, Y.-M.

中文

中华微生物学和免疫学杂志

0254-5101

2011

31

2

124-128

10.3760/cma.j.issn.0254-5101.2011.02.007

国家自然科学基金（30970165） 湖南省自然科学基金（09JJ3059）

本校为第一且通讯机构

目的 分析沙眼衣原体(Chlamydia trachomatis,Ct)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其生物学特征.方法 PCR扩增pORF5质粒蛋白编码基因,构建pORF5原核表达重组体并诱导表达融合蛋白;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;分析pORF5质粒蛋白在感染细胞中的分布及表达模式特征.采用ELISA分析pORF5质粒蛋白在自然感染状态下的表达情况及免疫原性.结果 pORF5主要分布于宿主细胞质,但也少量分布在原体(EB)和网状体(RB)上,在细胞质中的分布模式与衣原体分泌蛋白酶样活性因子(CPAF)...

Objective: To localize and characterize the plasmid protein p ORF5 in the Chlamydia trachomatis(Ct) infected cells. Methods: The open reading frame encoding for p ORF5 protein from the Ct plasmid was amplified and cloned into the p GEX-6p vector. The recombinant plasmid p GEX-p ORF5 was transformed into XL1-blue E. coli to express fusion protein with the glutathione-s-transferase (GST). After purified with Glutathione Sepharose 4B beads, the p ORF5 fusion protein was used to immunize mice to make monoclonal and polyclonal antibody. The antibodies were used to localize the endogenous p ORF5 pro...