论文部分内容阅读
目的 :构建东部马脑炎病毒E2基因的重组真核表达载体 ,对其DNA免疫原性进行观察。方法 :采用RT_PCR扩增东部马脑炎病毒全长E2基因 ,构建真核表达载体pcDNA_E2 ,以脂质体法转染COS7细胞 ,经蛋白印迹法和免疫荧光法证明E2基因可以表达后 ,用该重组质粒DNA免疫Balb c小鼠 ,并用免疫荧光法检测鼠血清中病毒的特异抗体。结果 :免疫印迹法、免疫荧光法检测表明E2基因在COS7细胞中获得瞬时表达 ,pcDNA_E2基因免疫小鼠可产生抗东部马脑炎病毒的特异抗体。结论 :东部马脑炎病毒E2基因的重组质粒DNA可刺激小鼠产生特异的抗东部马脑炎病毒体液免疫应答 ,为基因疫苗的研制奠定了基础
OBJECTIVE: To construct a recombinant eukaryotic expression vector for E2 gene of equine encephalitis in eastern part of China. The immunogenicity of DNA was observed. Methods: The full length E2 gene of eastern equine encephalitis was amplified by RT_PCR. The eukaryotic expression vector pcDNA_E2 was constructed and transfected into COS7 cells by lipofectamine 2000. After E2 gene was proved by Western blotting and immunofluorescence, Balb c mice were immunized with recombinant plasmid DNA and the specific antibodies against the virus in the serum were detected by immunofluorescence. Results: Western blotting and immunofluorescence assay showed that E2 gene was transiently expressed in COS7 cells. The pcDNA_E2 gene immunized mice could produce specific antibodies against equine encephalitis in the east. CONCLUSION: Recombinant plasmid DNA of equine encephalitis E2 E2 gene can stimulate mice to produce specific humoral immune response against eastern equine encephalitis virus, laying a foundation for the development of gene vaccines