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本研究根据GenBank登录的BJ01株SARS-CoV序列合成801bpS1基因片段,该片段被亚克隆至真核表达载体pcDNA3.1(+)得到重组质粒pcDNA3.1(+)/S1;转染Hela细胞,SDS-PAGE、Western-Blotting鉴定蛋白表达;肌注免疫BALB/c小鼠,利用ELISA法检测免疫后小鼠的抗SARS-CoVIgG及IFN-γ水平,MTT法检测T细胞增殖活性。结果显示,重组质粒pcDNA3.1(+)/S1可在Hela细胞内表达S1蛋白,免疫后小鼠的T细胞增殖活性增强,抗SARS-CoVIgG与IFN-γ水平升高。本实验说明pcDNA3.1(+)/S1可诱导小鼠产生一定的体液免疫和细胞免疫应答。
In this study, 801bpS1 gene fragment was synthesized based on the sequence of SARS-CoV BJ01 strain registered in GenBank. The fragment was subcloned into eukaryotic expression vector pcDNA3.1 (+) to obtain recombinant plasmid pcDNA3.1 (+) / S1. The expression of anti-SARS-CoVIgG and IFN-γ in the immunized mice was detected by ELISA. The proliferation activity of T cells was detected by MTT assay. The results showed that recombinant plasmid pcDNA3.1 (+) / S1 can express S1 protein in Hela cells, and the T cell proliferation activity and the level of anti-SARS-CoVIgG and IFN-γ in mice increased after immunization. This experiment shows that pcDNA3.1 (+) / S1 can induce certain humoral and cellular immune responses in mice.