沙眼衣原体CT058蛋白在感染细胞中的定位分析

Intracellular localization of the hypothetical protein CT058 in the Chlamydia trachomatis-infected cells

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。

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.