衣原体噬菌体phiCPGl衣壳蛋白Vpl对沙眼衣原体的作用研究  

The effect of chlamydiaphage phiCPG1 capsid protein Vpl on the Chlamydia trachomatis

作　　者：刘原君[1] 侯淑萍[1] 卫酒荣 李燕[1] 齐蔓莉[1] 王惠平[1] 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性传播疾病研究所,300052 [2]山西省阳城县凤城中心卫生院

出　　处：《中华微生物学和免疫学杂志》2012年第5期403-407,共5页Chinese Journal of Microbiology and Immunology

基　　金：国家自然科学基金（30671879,31100138）

摘　　要：目的将原核表达纯化的衣原体噬菌体phiCPGl衣壳蛋白Vpl作用于沙眼衣原体（Chlamydia trachomatis，Ct），以期发现该Vpl蛋白对Ct生长的影响。方法原核表达并纯化噬菌体phiCPGl的衣壳蛋白Vpl，将纯化后的蛋白复性后与ct的标准株或临床野生株（终浓度为53μg／m1）室温孵育3h后接种至单层致密McCoy细胞中，培养过程中均加入了Vpl蛋白，48h后碘染色包涵体计数和透射电镜观察Vpl蛋白对Ct生长的影响；MTT法检测Vpl蛋白对McCoy细胞系的细胞毒性作用；液体培养基稀释法检测Vpl蛋白对大肠杆菌BL21和DH5ct的抗菌作用。结果衣壳蛋白Vpl对Ct标准株E和D型的抑制率分别为78％和72％，对Ct临床野生株的抑制率为40％-70％，透射电镜结果显示Vpl蛋白能够抑制Ct的正常发育，使包涵体内出现异常增大的网状体。而该Vpl蛋白对非衣原体的其他生物体如大肠杆菌BL21、DH5α和真核细胞McCoy的生长没有影响。结论噬菌体衣壳蛋白Vpl对Ct的生长具有明显的抑制作用，为临床上Ct感染的治疗提供了新的思路。Objective To investigate the effect of recombinant chlamydiaphage phiCPG1 capsid protein Vpl on Chlamydia trachomatis（Ct） after Vpl was co-cultured with Ct （reference strains and clinical strains ）. Methods The recombinant chlamydiaphage phiCPG1 capsid protein Vpl was expressed and purified. Equal amount of Ct standard strains （E/UW-5/Cx and D/UW-3/Cx） or clinical strains, which had been incubated with Vpl protein at the concentration of 53 μg/ml for 3 h at room temperature, were inoculated into McCoy. After cell culture, idione stain and transmission electron microscope were used to observe the effect of Vpl on the Ct. The effect of Vpl protein on the cell line McCoy was determined by MTT assay, the responses of Escherichia coli BI21 and DH5α toward Vpl protein were determined using broth microdilu- tion assays. Results Vpl had obviously inhibitive effect on Ct, the inhibition ratios were about 40%-70% in clinical strains, 72% in reference strain D and 78% in E, respectively. Abnormally enlarged RBs were observed after Vpl-treatment and Vpl could arrest chlamydial developmental cycle using electron micro- scope. There was no effect of Vpl on McCoy cells or bacteria BL21 or DH5ot. Conclusion The recombinant Vpl from phiCPG1 has obviously inhibitive effect on the growth of Ct, it will be helpful for the treatment of Ct infection in clinic.

关 键 词：沙眼衣原体 衣原体噬菌体 VP1蛋白 抑制作用 

分 类 号：R759[医药卫生—皮肤病学与性病学]