辛德毕斯病毒复制子载体系统的构建  

Construction and Identification of Replicon Vector Derived from an Infectious Full-length cDNA Clone of a Sindbis Virus

作　　者：朱武洋[1] 付士红[1] 王力华[1] 何英[1] 唐青[1] 梁国栋[1] 

机构地区：[1]中国疾病预防控制中心病毒病预防控制所传染病预防与控制国家重点实验室,北京100052

出　　处：《病毒学报》2009年第2期143-147,共5页Chinese Journal of Virology

摘　　要：To construct vector system of XJ-160 virus,a Sindbis virus isolated in China,recombinant vector pBRepXJ together with its helper plasmid pBR-H were derived from XJ-160 viral infectious clone pBR-XJ160 by overlap-PCR.To quantitatively and qualitatively verify the function of the replicon system,recombinant plasmids pSinRep-EGFP,pBRepXJ-EGFP,pSinRep-R and pBRepXJ-R were constructed by cloning report genes of enhanced green fluorescent protein(EGFP) or Renilla luciferase(R.luc) into pBRepXJ or pSinRep5,a commercial Sindbis vector.And in Vitro-synthesized RNA from expression vectors were electroporated into BHK-21 cells.The results indicated that the replicon vector system was capable of self-replicating in host cell,and the expression efficiency of heterologous genes corresponded with that of the commercial Sindbis vector(pSinRep5).Our study laid the basis for developing alphavirus vector system with Chinese intellectual property.To construct vector system of XJ-160 virus, a Sindbis virus isolated in China, recombinant vector pBRepXJ together with its helper plasmid pBR-H were derived from XJ-160 viral infectious clone pBR- XJ160 by overlap-PCR. To quantitatively and qualitatively verify the function of the replicon system, recombinant plasmids pSinRep-EGFP, pBRepXJ-EGFP, pSinRep-R and pBRepXJ-R were constructed by cloning report genes of enhanced green fluorescent protein （EGFP） or Renilla luciferase （R. luc） into pBRepXJ or pSinRepS, a commercial Sindbis vector. And in Vitro-synthesized RNA from expression vectors were electroporated into BHK-21 cells. The results indicated that the replicon vector system was capable of self-replicating in host cell, and the expression efficiency of heterologous genes corresponded with that of the commercial Sindbis vector （pSinRep5）. Our study laid the basis for developing alphavirus vector system with Chinese intellectual property.

关 键 词：辛德毕斯病毒 XJ-160病毒 复制子载体 

分 类 号：R373.3[医药卫生—病原生物学]