东部马脑炎病毒E2基因的表达及DNA免疫的初步研究

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，用庐重组质粒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性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。

Expression and immunogenicity of the E2 gene of eastern equine encephalomyelitis virus

Objective:To investigate the expression and immunogenicity of E2 gene DNA of eastern equine encephalomyelitis virus.Methods:The E2 gene of eastern equine encephalomyelitis virus was amplified by RT_PCR and then cloned into eukaryotic expression vector pcDNA3.1. The recombinant was transfected into COS7 cells by lipofectin. Western blotting and immunofluorescence were used to identify the expression of the E2 gene. After Balb/c mice were vaccinated with pcDNA_E2,immunofluorescence was used to detect the antibody titers of immunized mice sera.Results:Western blotting test and immunofluorescence showed that E2 gene could be expressed in COS7 cells. pcDNA_E2 induced specific antibody against EEEV in immunized mice.Conclusion:The recombinant eukaryotic expression vector pcDNA_E2 was capable of stimulating specific humoral immunity against EEEV in Balb/c mice.