SARS-CoV S蛋白表位的表达及鉴定  

Preparation and characterization of the epitopes of SARS coronavirus spike protein

作　　者：房凯[1] 钟连声[1] 马汝海[2] 王绍成[1] 潘忠诚[1] 张玉魁[1] 何群[1] 赵雨杰[1] 

机构地区：[1]中国医科大学基础医学院生物信息学教研室,辽宁沈阳110001 [2]中国医科大学基础医学院化学教研室,辽宁沈阳110001

出　　处：《细胞与分子免疫学杂志》2007年第11期1007-1009,共3页Chinese Journal of Cellular and Molecular Immunology

基　　金：辽宁省科技厅科技计划资助项目(2004225009)

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。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 Ni^2＋ 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 SARSCoV.

关 键 词：SARS冠状病毒 S蛋白 表位 抗原 

分 类 号：R373[医药卫生—病原生物学]