中华传染病杂志
中華傳染病雜誌
중화전염병잡지
CHINESE JOURNAL OF INFECTIOUS DISEASES
2012年
10期
583-586
,共4页
刘原君%孙毅娜%姚卫锋%李燕%李卓然%卫酒荣%刘全忠
劉原君%孫毅娜%姚衛鋒%李燕%李卓然%衛酒榮%劉全忠
류원군%손의나%요위봉%리연%리탁연%위주영%류전충
衣原体,沙眼%细菌噬菌体%病毒壳体%细菌外膜蛋白质类
衣原體,沙眼%細菌噬菌體%病毒殼體%細菌外膜蛋白質類
의원체,사안%세균서균체%병독각체%세균외막단백질류
Chlamydia trachomatis%Bacteriophages%Capsid%Bacterial outer membrane proteins
目的 发现噬菌体phiCPG1衣壳蛋白 Vp1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phiCPG1的衣壳蛋白Vp1,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Vp1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Vp1;抗Vp1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Vp1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Vp1能特异结合D型沙眼衣原体标准株的外膜蛋白rPmpI.结论 沙眼农原体外膜中有Vp1的结合位点,可能通过和rPmpI的结合从而特异性地与沙眼农原体结合.
目的 髮現噬菌體phiCPG1衣殼蛋白 Vp1在沙眼衣原體中的結閤位點,為理解噬菌體與衣原體的相互作用提供線索.方法 誘導錶達噬菌體phiCPG1的衣殼蛋白Vp1,通過膠迴收進行目的蛋白的純化,複性後以Far-Western印跡技術檢測Vp1是否能與衣原體外膜中重組多形外膜蛋白(rPmp)傢族及主要外膜蛋白(MOMP)結閤.結果 在大腸埃希菌中成功錶達蛋白Vp1;抗Vp1的單剋隆抗體作為一抗進行Western印跡,結果無特異性條帶齣現,可見針對Vp1的單剋隆抗體不與任何一種rPmp結閤.Far-Western印跡結果顯示,在rPmpI的位置齣現明顯的條帶,在相對于其他rPmp和MOMP的位置無特異性條帶的齣現,證實Vp1能特異結閤D型沙眼衣原體標準株的外膜蛋白rPmpI.結論 沙眼農原體外膜中有Vp1的結閤位點,可能通過和rPmpI的結閤從而特異性地與沙眼農原體結閤.
목적 발현서균체phiCPG1의각단백 Vp1재사안의원체중적결합위점,위리해서균체여의원체적상호작용제공선색.방법 유도표체서균체phiCPG1적의각단백Vp1,통과효회수진행목적단백적순화,복성후이Far-Western인적기술검측Vp1시부능여의원체외막중중조다형외막단백(rPmp)가족급주요외막단백(MOMP)결합.결과 재대장애희균중성공표체단백Vp1;항Vp1적단극륭항체작위일항진행Western인적,결과무특이성조대출현,가견침대Vp1적단극륭항체불여임하일충rPmp결합.Far-Western인적결과현시,재rPmpI적위치출현명현적조대,재상대우기타rPmp화MOMP적위치무특이성조대적출현,증실Vp1능특이결합D형사안의원체표준주적외막단백rPmpI.결론 사안농원체외막중유Vp1적결합위점,가능통과화rPmpI적결합종이특이성지여사안농원체결합.
Objective To investigate the binding protein of chlamydiaphage phiCPG1 capsid protein Vp1 on chlamydia trachomatis outer membrane.Methods The bacterium with recombinant plasmid Vp1/pet30a( + ) was induced.The expressed protein was purified by gel recycling.FarWestern blot was utilized to' investigate the binding protein of Vp1 on chlamydial outer membrane,including recombinant polymorphic outer membrane protein (rPmp) and major outer membrane protein (MOMP).Results The recombinant protein Vp1 was successfully expressed in E.coli.Monoclonal antibody against Vp1 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 Vp1 could specifically bind with rPmpI protein on the chlamydial outer membrane of serotype D.Conclusions There is a binding site of Vp1 on the chlamydia trachomatis outer membrane.Vp1 may play an important role in the interaction between the chlamydiaphage and the chlamydiae.
