作　　者： ; ; ;

机构地区： 淮阴工学院生命科学与化学工程学院

出　　处： 《江苏农业学报》 2011年第6期1216-1221,共6页

摘　　要： 编码SARS冠状病毒S蛋白(Spike protein)的C端序列S2较为保守,可作为SARS病毒疫苗研究的候选抗原基因。该研究拟构建基于SARS病毒S2蛋白的重组腺病毒疫苗并对其进行免疫学初步分析。首先通过非对称PCR方法,将人工合成的S2蛋白基因的两个片段f3和f4进行拼接得到完整的S2片段,利用Ad5腺病毒系统构建S2蛋白基因的重组腺病毒并在AD293细胞中包装成病毒颗粒,采用间接免疫荧光和蛋白免疫印迹法对S2蛋白基因的表达进行鉴定,然后用此重组腺病毒免疫小鼠对其抗体生长情况进行分析。免疫荧光和蛋白印迹分析结果显示,携带SARS病毒S2蛋白基因的重组腺病毒rAd-S2能在细胞中成功表达,表达的S2蛋白能被马抗SARS血清所识别。ELISA分析结果表明,用表达SARS病毒S2蛋白的重组腺病毒rAd-S2免疫小鼠,能有效刺激小鼠机体产生抗体,重组腺病毒免疫小鼠后所产生的抗体水平最高能达到104以上。可见通过对SARS病毒S2蛋白基因重组腺病毒的成功构建与免疫学初步分析,能为SARS疫苗研究奠定基础。 S2 protein,the carboxyl terminus of SARS spike glycoprotein,is a conserved protein and can be a candidate antigenic gene for the SARS vaccine research.The aim of this study is to construct recombinant adenovirus based on S2 protein and investigate its immune effects.The S2 gene of SARS spike glycoprotein was synthesized by asymmetric PCR.Ad5 recombinant adenovirus system was used as a vector to express S2 protein and the recombinant adenovirus was verified by Western blotting and IFA methods.BALB/c mice were immunized with two doses of recombinant adenovirus through intraperitoneal route and ELISA was applied to detect its serum antibody titers.Results showed that the recombinant adenovirus（rAd-S2） expressed S2 protein successfully and the S2 protein could be recognized by horse anti-SARS sera.Mice immunized with rAd-S2 elicited high titers of antibodies against SARS coronavirus,the highest antibody titer being up to 104.Through successful construction of recombinant adenovirus of SARS S2 protein,it laid strong foundation for the vaccine research on SARS.

关 键 词： 刺突蛋白 蛋白 重组腺病毒 免疫

领　　域： [生物学]