衣原体噬菌体phiCPG1衣壳蛋白Vp1在D型沙眼衣原体外膜中结合位点的检测  

The detection of the binding protein of chlamydiaphage phiCPG1 capsid protein Vpl on chlamydial outer membrane of serotype D

作　　者：刘原君[1] 孙毅娜[2] 姚卫锋[1] 李燕[1] 李卓然[1] 卫酒荣 刘全忠[1] 

机构地区：[1]天津医科大学总医院皮肤性病科天津市性传播疾病研究所,300052 [2]天津医科大学代谢病医院内分泌研究所 [3]山西省阳城县凤城中心卫生院

出　　处：《中华传染病杂志》2012年第10期583-586,共4页Chinese Journal of Infectious Diseases

基　　金：国家自然科学基金资助项目（31100138、30901460、30671879）

摘　　要：目的发现噬菌体phiCPGl衣壳蛋白Vpl在沙眼衣原体中的结合位点，为理解噬菌体与衣原体的相互作用提供线索。方法诱导表达噬菌体phiCPGl的衣壳蛋白Vpl，通过胶回收进行目的蛋白的纯化，复性后以Far-Western印迹技术检测Vpl是否能与衣原体外膜中重组多形外膜蛋白（rPmp）家族及主要外膜蛋白（MOMP）结合。结果在大肠埃希菌中成功表达蛋白Vpl；抗Vpl的单克隆抗体作为一抗进行Western印迹，结果无特异性条带出现，可见针对Vpl的单克隆抗体不与任何一种rPmp结合。Far-Western印迹结果显示，在rPmpI的位置出现明显的条带，在相对于其他rPmp和MOMP的位置无特异性条带的出现，证实Vpl能特异结合D型沙眼衣原体标准株的外膜蛋白rPmpI。结论沙眼衣原体外膜中有Vpl的结合位点，可能通过和rPmpI的结合从而特异性地与沙眼衣原体结合。Objective To investigate the binding protein of chlamydiaphage phiCPG1 capsid protein Vpl on chlamydia trachomatis outer membrane. Methods The bacterium with recombinant plasmid Vpl/pet30a（ ＋ ） was induced. The expressed protein was purified by gel recycling. Far- Western blot was utilized to investigate the binding protein of Vpl on chlamydial outer membrane, including recombinant polymorphic outer membrane protein （rPmp） and major outer membrane protein （MOMP）. Results The recombinant protein Vpl was successfully expressed in E. colL Monoclonal antibody against Vpl was used as primary antibody in Western blot, and no specific band was present, which indicated that the monoclonal antibody did not specifically bind with any rPmp. Far-Western blot results showed that there was an obvious band for the rPmpI, but no specific band for other rPmp and MOMP, which suggested that Vpl could specifically bind with rPmpI protein on the chlamydial outer membrane of serotype D. Conclusions There is a binding site of Vpl on the chlamydia trachomatis outer membrane. Vpl may play an important role in the interaction between the chlamydiaphage and the chlamydiae.

关 键 词：衣原体 沙眼 细菌噬菌体 病毒壳体 细菌外膜蛋白质类 

分 类 号：R374[医药卫生—病原生物学]