我国D2-43病毒株PrM-E基因的复制型载体质粒DNA的免疫原性(英文)  

The Immunogenicity of Replicative Virus Plasmid DNA Containing PrM-E Gene of Chinese D2-43 Virus Strain

作　　者：陈水平[1] 秦鄂德[1] 于曼[1] 胡志君[1] 赵卫[1] 范宝昌[1] 王鹏程[1] 杨佩英[1] 

机构地区：[1]军事医学科学院微生物流行病研究所,北京100071

出　　处：《中国生物化学与分子生物学报》2002年第2期151-155,共5页Chinese Journal of Biochemistry and Molecular Biology

基　　金：国家自然科学基金资助 (No .39770 0 36 )~~

摘　　要：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 study the immunogenicity of replicative plasmid DNA containing PrM-E gene of Chinese D2-43 virus strain and lay a foundation for new vaccine of dengue, the PrM-E gene was cut from pGEM-T-ME with ApaⅠ and ClaⅠ and inserted into replicative semliki forest virus(SFV) vector. The recombinant plasmid DNA was electroporated into BHK21 cells, then expression products specific to dengue virus type 2 could be detected by IFA (indirect immunofluorescence assay) in transfected cells. Endotoxin-free plasmid DNA was prepared with EndoFree plasmid Maxi kit. Mice were immunized with different doses of the recombinant SFV plasmid DNA intramuscularly at multiple sites. The titres of antibody to dengue virus type 2 was measured by IFA. And sera with recombinant plasmid, could react with antigen of dengue 2 virus on microscope slides.The results showed that recombinant SFV plasmid DNA containing the PrM-E gene of D2-43 virus strain, could produce specific antibody to dengue 2 virus in mice, but the titre was low.

关 键 词：D2-43病毒株 PrM-E基因 复制型载体质粒DNA 免疫原性 登革病毒 DNA免疫 

分 类 号：R373.33[医药卫生—病原生物学]