摘要
目的构建A型流感病毒M基因DNA疫苗质粒载体并对免疫后小鼠体内免疫应答反应进行分析和评价。方法将H3N2亚型流感病毒M基因通过RT-PCR扩增后克隆构建DNA疫苗载体pBudCE4.1-M,并将pBudCE4.1-M质粒体外瞬时转染COS7细胞并通过Western blot鉴定M1蛋白的表达,然后将pBudCE4.1-M质粒免疫小鼠,通过ELISA对小鼠体内生成的M1蛋白lgG抗体进行鉴定。结果 M基因被成功克隆到质粒载体pBudCE4.1上,并在体外COS7细胞中表达出28kD的M1蛋白。疫苗质粒免疫小鼠后4周,在小鼠血清中检测到M1蛋白lgG抗体的吸光度有明显升高。结论成功构建了一种A型流感病毒M基因DNA疫苗载体,将其免疫小鼠后成功诱导小鼠体内产生了特异性的lgG抗体。
Aim To develop M gene DNA vaccine vectors of influenza A virns and evaluate its immune response in vivo. Methods Amplified by RT-PCR,Mgene （H3N2）were cloned into vector pBudCE4.1. Then the reconrbinant plasmid pBudCE4.1-M was transfected transiently into COS7 cells and the expression of M1 protein was tested by western blot. Mice were immunized with the plasmid pBudCE4.1-M and the Ml-specific antibody was detected by ELISA. Results The M gene was cloned into plasmid pBudCE4.1 successfully and the 28kD M1 proteins were detected in the transfected COS7 cells. The absorbance value of M1-specific antibody increased distinctly in serum of the immunized mice. Conclusion A DNA vaccine plasmid pBudCE4.1-M was developed successfully and the Ml-specific antibody was also induced in the mice immunized with the pBudCE4.1-M plasmid.
出处
《中国热带医学》
CAS
2011年第7期800-802,共3页
China Tropical Medicine
