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目的构建SARS病毒核衣壳蛋白(N)的真核表达质粒,并研究其在小鼠体内诱导的体液免疫应答。方法采用PCR方法体外扩增SARS病毒N蛋白基因片段,定向克隆入真核表达载体pVAC,构建pVAC-N重组质粒;大量制备该重组质粒,经基因枪腹部皮内注射免疫BALB/c小鼠三次,间接ELISA法测定免疫鼠IgG抗体效价。结果成功构建了重组表达质粒pVAC-N,免疫小鼠后可诱导产生出特异性的IgG抗体。结论构建的真核表达质粒pVAC-N能诱导小鼠产生特异性抗体,为SARS疫苗的进一步研究打下基础。
Objective To construct eukaryotic expression plasmid of nucleocapsid protein (N) of SARS virus and study its humoral immune response induced in mice. Methods The gene fragment of SARS-CoV N protein was amplified by PCR in vitro and cloned into the eukaryotic expression vector pVAC to construct pVAC-N recombinant plasmid. The recombinant plasmid was prepared in large quantities and BALB / c mice were immunized intradermally with gene gun Indirect ELISA was used to determine the titer of immunized rat IgG antibody. Results The recombinant plasmid pVAC-N was constructed successfully. The immunized mice were induced to produce specific IgG antibodies. Conclusion The constructed eukaryotic expression plasmid pVAC-N can induce the production of specific antibodies in mice and lay the foundation for the further study of SARS vaccine.