【摘要】 目的 初步研究SARS-Coy病毒M蛋白基因的真核表达效率和免疫原性为进一步的疫苗研究奠定基 础。方法 以PCR方法在全病毒基因组文库中获得全长M基因,分别克隆至pcDNA4-his／myc和pVAON33载 体获得重组质粒pcDNA4-his／myc-M和pVAON33-M。以Western Blot方法利用融合分子羧基段的myc表位检 测pcDNA4-his／myc-M转染的293T细胞中M蛋白的真核表达。以ELISA方法检测pVAON33-M质粒基因免 疫小鼠诱导产生特异性抗血清。结果pcDNA4-his／myc-M转染后48 h可检测到SARS-Cov M蛋白表达。 pVAON33-M基因免疫能够诱导产生M特异的体液免疫应答。结论 本研究的结果表明SARS-Cov M蛋白可 以在真核细胞中有效表达并有良好免疫原性,可以作为SARS-Cov疫苗研制的候选抗原之一。更多还原

【Abstract】 Purpose To investigate the eukaryotic expression and immunogenecity of SARS-Cov M protein. Methods RT-PCR based gene cloning strategy was applied to get the whole SARS-Cov M gene from the genome of SARS-Cov virus. The M gene fragment was then cloned into pcDNA4-his/myc and pVAON33 vectors. Western Blot was employed for the detection of the M-his/myc fusion protein and ELISA was used to trace specific humoral immune response elicited by the pVAON33-M gene immunization. Results After transfection, M protein was proved to express well. Subsequent immuniza-tion test proved that specific anti-M antibody was elicited in the BALB/c mice. Conclusions Our results suggested that M protein was a candidate target for vaccine development.更多还原