2型猪链球菌胞壁蛋白MRP的截短表达、纯化及抗原保护性评价  

Expression,purification and protective antigen analysis of cell wall protein MRP of Streptococcus suis type 2

作　　者：王萍萍[1,2] 骈亚亚[2] 袁媛[2] 郑玉玲[2] 姜永强[2] 熊正英[1] 

机构地区：[1]陕西师范大学体育学院,陕西西安710026 [2]军事医学科学院微生物流行病研究所病原微生物生物安全国家重点实验室,北京100071

出　　处：《细胞与分子免疫学杂志》2012年第2期117-119,共3页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金资助项目(30870091)

摘　　要：目的:扩增猪链球菌2型(Streptococcus suis 2)05Z33中的mrp基因,并在大肠杆菌BL21中重组表达,以获得高纯度的重组MRP截短蛋白,验证其抗原保护性。方法:以05ZYH33基因组为模板并设计引物扩增目的片段,构建表达质粒,利用亲和层析对目的蛋白进行纯化,免疫新西兰大白兔制备抗血清并测其效价。通过抗原保护实验及抗血清调理的全血杀伤实验验证重组MRP蛋白抗原保护性。结果:重组MRP蛋白在大肠杆菌中高浓度表达,纯化获得了高纯度的MRP截短蛋白并制备获得了抗血清。经该蛋白免疫过的CD-1小鼠与阴性对照组相比,存活率显著升高。经抗血清调理后,野生株05ZYH33在全血中的存活率显著降低,突变株ΔMRP无显著变化。结论:MRP蛋白有较高的抗原保护性。AIM： To amplify the mrp gene of Streptococcus suis type 2 05ZYH33,express it in E.coli BL21 in order to acquire high purity recombinant protein MRP,then evaluate the protective antigen of recombinant protein MRP.METHODS： Using PCR technology to obtain the product of mrp gene of 05ZYH33,and then cloned it into the expression vector pET28a（＋）.The recombinant protein was purified by affinity chromatography,later immunized New Zealand rabbit to gain anti-serum,then test the anti-serum titer by ELISA.The opsonophagocytic killing test demonstrated the abilities of protective antigen of MRP.RESULTS： The truncated of MRP recombinant protein in E.coli BL21 expressed by inclusion bodies,and purified it in high purity.After immunoprotection,the survival condition of CD-1 was significantly elevated.The survival rate of wild-type strain 05ZYH33 in blood was apparently decreased after anti-serum opsonophagocyticed,but the mutant ΔMRP showed no differences.CONCLUSION： MRP represent an important protective antigen activity.

关 键 词：猪链球菌2型 MRP 抗原保护性 

分 类 号：Q936[生物学—微生物学]