Construction of Replicon Expression Vector Derived from XJ-160 Virus

Abstract：XJ一1 60 is the first Chinese isolate of a Sindbis—like virus，an d its genome was completely sequenced in our previous work．In this investigation，a full—length genomic cDNA clone Was firstly constructed from RT—PCR products of the viral RNA．Then．the plasmid of replicon expression vector (pRepxj 1 60)was derived from this cDNA clone replacin~viral structural sequence with multiclonal seq uence by DNA recombination technique．To verify function of this vector,reporter genes．EGFP an d cZ．were cloned into this plasmid．respectively．And RNA tran scripted from expression plasmid Was inducted into BHK一2 l cell line． resulting in expression of green fluorescent protein an d 13 ．galactosidase 14 hours later．Th e results indicated mat the replicon vector derived from )(J—l60 virus was self-replicating and that the following gene expression was efficient．Our study settled the basis for developing all：Ihavirus vector system with Chinese intellectual property．