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观察含我国登革 2型病毒株 (D2 4 3)的PrM E基因的复制型SFV(semlikiforestvirus)重组质粒DNA的免疫原性 ,为登革新型疫苗的研制提供依据 .将PrM E基因自T载体上切下 ,插入复制型SFV病毒载体质粒DNA中 .将此重组质粒DNA以电穿孔法导入BHK2 1细胞 ,用间接免疫荧光法在感染细胞内可检测到登革 2型病毒特异蛋白的表达 .采用去除内毒素的质粒提取试剂盒制备重组质粒DNA ,然后以不同剂量通过肌肉多点注射途径免疫Balb c鼠 ,获得的鼠血清可与登革D2 4 3感染的C6 36抗原片起特异的抗原抗体反应 .结果表明 ,含登革 2型病毒PrM E基因的复制型SFV病毒载体质粒DNA在Balb c鼠中可诱导登革 2型病毒特异抗体的产生 ,但抗体水平较低 .
To observe the immunogenicity of the recombinant plasmid DNA of the replicative SFV (semlikiforestvirus) containing the PrM E gene of Dengue 2 type strain (D2 4 3) in China, and to provide a basis for the development of a new dengue vaccine.The PrM E gene was cloned from T vector And inserted into the plasmid DNA of the replicative SFV virus vector.The recombinant plasmid DNA was introduced into BHK2 1 cells by electroporation and the expression of Dengue virus type 2 protein was detected by indirect immunofluorescence in infected cells. Recombinant plasmid DNA was prepared by removing endotoxin plasmid extraction kit, and then Balb C mice were immunized by multi-point injection at different doses. The obtained mouse serum could be used as a specific antigens against the C6 36 antigen plate infected with dengue D2 43 The results showed that the plasmid DNA of replicative SFV virus vector containing PrM E gene of dengue 2 could induce the production of Dengue virus type 2 specific antibody in Balb c mice, but the antibody level was low.