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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. 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 were co-populated that of the commercial Sindbis vector (pSinRep5) .Our study laid the basis for developing alphavirus vector system with Chinese intellectual property.