摘要
目的 在本室前期工作的基础上 ,进一步进行汉坦病毒 S基因 0 .7kb片段的真核表达及基因免疫的研究 .方法 构建汉坦病毒 76 - 118株 S基因 5 端 70 0 bp片段的真核表达载体 pc DNA3.1- S0 .7,脂质体法转染 COS- 7细胞 ,免疫荧光检测表达结果 ;用该质粒免疫 BAL B/c小鼠 ,并用免疫荧光法及 EL ISA法检测基因免疫的体液免疫应答效果 .结果 免疫荧光检测结果表明 ,目的基因在 COS- 7细胞中获得瞬时表达 ;用 pc DNA3.1- S0 .7基因免疫小鼠可引起抗汉坦病毒核蛋白 (NP)特异性的抗体应答 .结论 汉坦病毒 S基因 0 .7kb片段可刺激机体产生特异的抗汉坦病毒体液免疫应答 ,为进一步进行细胞免疫反应的检测及基因疫苗的研究提供实验基础 .
AIM To investigate the expression and immunogenicity of recombinant eukaryotic plasmid containing competent fragment of Hantaan virus S segment. METHODS Recombinant eukaryotic expression vector pcDNA3.1 S0.7 was constructed by cloning S segment 0.7 kb fragment into pcDNA3.1 (+) and transfected COS 7 cells by lipofectin.Immunoflourescence was used to identify the expression. After BALB/c mice were vaccinated by pcDNA3.1 S0.7, immunoflourescence and ELISA were used to detect the antibody titers of immunized mice sera. RESULTS Eukaryotic expression vector pcDNA3.1 S0.7 was proved to be successfully constructed by restriction enzyme analysis. Immuno flourescence and ELISA results showed that the truncated NP was transient expressed in COS 7 cells and pcDNA3.1 S0.7 induced specific antibody against NP in immunized mice. CONCLUSION It is suggested that the 0.7 kb competent fragment of hantaan virus S segment could directly stimulate specific anti Hantaan virus humor immunity in BALB/c mice.
出处
《第四军医大学学报》
北大核心
2001年第14期1287-1289,共3页
Journal of the Fourth Military Medical University
基金
国家自然科学基金资助项目 ( 3 0 0 70 686
3 9970 70 2 )
国家教育部骨干教师资助计划资助项目
