【摘要】 目的:制备SARS冠状病毒S蛋白串联表位重组蛋白,为SARS的防治提供新型抗原蛋白。方法:应用抗原表位分析软件分析S蛋白的表位。选取其中16个表位,设计并合成表位串联重组蛋白编码基因Z,构建其原核细胞表达重组体,在大肠杆菌BL21(DE3)表达该重组表位蛋白Z,应用Ni离子亲合层析法纯化重组蛋白Z做抗原,采用皮下注射法免疫新西兰白兔。斑点杂交法测定抗Z-血清中S蛋白的抗体,ELISA法检测Z蛋白的抗原性。结果:构建了S蛋白重组表位蛋白Z的原核表达体,在BL21菌中表达了Z蛋白,Z蛋白免疫新西兰白兔后获得了抗Z血清。斑点杂交分析显示,抗Z血清识别哺乳动物细胞中表达的S蛋白。ELISA检测结果显示,抗Z血清识别SARS冠状病毒抗原。结论:建立了制备SARS冠状病毒S蛋白重组表位蛋白的方法,为防治SARS提供了新型抗原蛋白Z。更多还原

【Abstract】 AIM:To prepare the recombinant epitopes of SARS-CoV spike protein and study their antigenic property to spike protein.METHODS:The epitopes of SARS-CoV spike protein were analyzed by epitope prediction software.A novel gene,named Z,encoding 16 predicted epitopes of spike protein was designed and synthesized using chemical method.Z gene was cloned into vector pET-28a(+),expressed in E.coli BL21(DE3) and purified by Ni2+ affinity method.Z protein was used as antigen to immunize the rabbit and anti-Z sera was collected.Then the antigenic property of Z protein to SARS-CoV spike protein was analyzed by dot-blot and ELISA assay.RESULTS:Z gene was successfully designed and expressed in E.coli BL21(DE3).Dot blot analysis showed that SARS-CoV spike protein,which was expressed and purified from mammal cells,can be detected by anti-Z sera from rabbit.ELISA analysis indicated the SARS-CoV antigen prepared from SARS-CoV lysates can be detected by anti-Z sera.CONCLUSION:The predicted epitopes of Z protein can induce the development of SARS-CoV spike protein antibody in rabbits,which provides a new protein for developing vaccine against SARS-CoV.更多还原