SARS相关冠状病毒M蛋白基因重组核酸疫苗的实验研究

目的：构建针对SAILS相关冠状病毒M蛋白基因的重组核酸疫苗，观察其免疫小鼠后肌体的抗体产生情况，探讨其作为抗SAILS病毒疫苗的可能性。方法：通过分子生物学的方法构建SARS相关冠状病毒M蛋白基因真核表达质粒pcDNA3．1／M。经肌注免疫健康BALB／c小鼠，在免疫后的2．4、6周取小鼠血清，用ELISA法检测其中的抗体。结果：接种含M基因的真核表达质粒pcDNA3．1／M的低剂量组和高剂量组的小鼠血清在接种2周后就可检测出SARS-CoV特异性IgG，第4周时，这种特异性IgG水平有升高趋势，第6周时，血清中的抗体含量与4周时无大的差异。高剂量组产生抗体与低剂量组产生抗体无显著差异。pcDNA3．1空载体接种的对照组未检测出特异性抗体（其OD值低于0．18）。结论：构建的真核表达质粒pcDNA3．1／M能诱导小鼠产生了针对SARS的抗体。

Experiment study of recombinant DNA vaccine against SARS coronavirus M gene

Objective:To construct recombinant DNA vaccine containing SARS-CoV M gene and to observe the production of SARS-CoV specific antibodies in mice. Methods: A recombinant plasmid-pcDNA3. 1/M was constructed by molecular biology technique. The mice was immunized with the recombinant DNA vaccine. After 2,4,6 week,the SARS-CoV antibodies in the sera of mice were detected with ELISA kit. Results: After 2 weeks,SARS-CoV specific antibodies were detected in mice. There were an ascended trend in antibodies concentration at 4 weeks. And simultanity,SARS-CoV specific antibodies was not detected in mice injected with vacancy carrier. There were not remarkable difference in mice injected with high and low quantity plasmid. Conclusion: A recombinant plasmid pcDNA3. 1/M could induce SARS-CoV specific antibodies in mice.