摘要
目的 :构建我国登革 2型病毒 4 3株prM E基因的甲病毒 (Semlikiforestvirus ,SFV)重组RNA ,为登革新型疫苗的研究奠定基础。方法 :首先将含有多种稀有限制酶位点的接头插入pSFV载体的多克隆位点 ,再把prM E基因克隆至该载体中。将获得的重组质粒DNA经体外转录后 ,通过电穿孔法将其转录的RNA产物导入宿主细胞 ,并采用间接免疫荧光法检测prM E基因的表达。结果 :已获得含多种酶位点的pSFV病毒载体 ,并且所构建的含prM E基因的重组pSFV病毒RNA ,在宿主细胞中的表达产物可与登革 2型病毒特异抗体起反应。结论 :构建的含prM E基因的重组pSFV病毒RNA可表达我国登革 2型病毒株的特异蛋白。
Objective: To construct the recombinant SFV RNA containing the prM E gene of the Chinese dengue 2 virus and to lay a foundation for development of new vaccines for dengue. Methods: First, a linker, which contains several rare restricted enzyme sites, was inserted into the multiple cloned sites of the pSFV1 vector. Then, the prM E gene was cloned into the modified vector. The linearized recombinant plasmid DNA was transcribed in vitro and the resulting RNA transcripts were introduced to BHK cells by electroporation. The expressed products were detected by indirect immunofluorescence technique. Results: The expressed products of the recombinant RNA could react with the specific antibody against the Chinese dengue 2 virus. Conclusions: The recombinant pSFV RNA which expresses the specific proteins of the Chinese dengue 2 virus was constructed. [
出处
《军事医学科学院院刊》
CSCD
北大核心
2000年第4期255-258,共4页
Bulletin of the Academy of Military Medical Sciences
基金
国家自然科学基金!资助项目 (39770 0 36 )
