金黄色葡萄球菌聚集因子A蛋白的原核表达及抗血清活性分析  

Prokaryotic expression of Staphylococcus aureus ClfA and assay for its antisera bioactivity

作　　者：陈智[1] 刘少宁[1] 牟凯[1] 曹丙蕾[1] 赵宏坤[1] 

机构地区：[1]山东农业大学动物科技学院,山东泰安271018

出　　处：《中国兽医学报》2010年第1期29-31,46,共4页Chinese Journal of Veterinary Science

基　　金：教育部高校科技创新工程重大项目培育资金资助项目(CNO706039)

摘　　要：采用PCR方法对金黄色葡萄球菌山东分离株ZFB1的聚集因子A(Clumping factor A,ClfA)基因进行扩增,并将其克隆到原核表达载体pGEX-6p-1中获得重组克隆pGEX/ClfA,使ClfA与GST进行融合表达,然后利用原核表达的蛋白免疫小鼠制备抗血清,并采用黏附与黏附抑制试验对抗血清的活性进行分析。在SDS-PAGE和Western-blot试验中,表达产物相对分子质量(约67 000)同预期结果相符,与兔源抗ZFB1全价血清有特异的抗体结合活性。在黏附与黏附抑制试验中发现ClfA抗体能够抑制金黄色葡萄球菌在细胞表面的黏附。结果表明,ClfA可以作为保护性抗原,有助于动物抵抗金黄色葡萄球菌的侵入、定植和感染,从而为奶牛金黄色葡萄球菌性乳腺炎的防治提供了科学的试验依据。Gene of clumping factor A （CIfA） from Staphylococcus aureus （S. aureus）Shandong isolate ZFB1 amplified by polymerase chain reaction （PCR） was cloned into pGEX-6p-1 getting the recombinant pGEX/ClfA,and the ClfA was fused and coexpressed with GST. In order to get the antisera of ClfA,mice were immunized with the expressed protein,meanwhile, the bioactivity of antisera was detected by adherence inhibition assay. The SDS-PAGE and West- ern-blot analysis indicated that the fusion protein was 67 000 in molecular weight and had immunological activity. The antisera of expressed protein can inhibit adherence activity of S. aureus to cell surface by adherence inhibition assay. As protective antigen,ClfA protein can contribute to protect animals from the invasion,planting and infection. The study provides a theoretical base for the prevention of bovine S. aureus mastitis.

关 键 词：金黄色葡萄球菌 乳腺炎 聚集因子A 原核表达 抗血清活性 

分 类 号：S852.61[农业科学—基础兽医学]