【摘要】 分析沙眼衣原体CT058蛋白在感染细胞中的定位。克隆表达CT058蛋白;纯化的CT058融合蛋白免疫小鼠制备多克隆抗体;间接免疫荧光法对CT058蛋白在沙眼衣原体感染细胞中的定位进行分析;Western blot检测CT058蛋白在原体和网状体中的表达情况。间接免疫荧光染色实验显示CT058蛋白位于包涵体内;鼠抗GST-CT058抗体与GST-CT058融合蛋白吸附后特异性染色消失,而与GST-CT232融合蛋白吸附后仍然可见GST-CT058抗体的包涵体染色特征;Western blot证实CT058蛋白在纯化的原体和网状体上均有表达。CT058蛋白定位于沙眼衣原体感染细胞的包涵体内。更多还原

【Abstract】 To tract the intracellular localization of the hypothetical protein CT058 in the Chlamydia trachomatis-infected cells.The gene ct058 was expressed as GST-CT058 fusion protein in E.coli and its antibody was prepared by immunizing mouse with the fusion protein.Then,the anti-GST-CT058 an-tibody was used to localize the endogenous protein in cells infected by C.trachomatis serovar L2 using indirect immunofluorescence assay(IFA).Western blot was carried out to detect whether CT058 pre-sents in the purified elementary body(EB) or reticulate body(RB).The anti-GST-CT058 antibody de-tected an inclusion-inside signal in an IFA.The intra-inclusion signal could be removed by pre-absorption of the antibody with the GST-CT058 fusion protein but not control GST-CT232.Fur-thermore,the mouse anti-CT058 antibody recognized the endogenous CT058 protein from purified EBs and RBs in a Western blot assay.The results of the current study support the general consensus that the hypothetical protein CT058 was localized within the inclusion body of the C.trachomatis-infected cells.更多还原