摘要：提取马立克氏病毒I型疫苗毒株CVI988的总DNA为模板，利用PCR一技术扩增出病毒生长非必需的US2 基因并克隆入T-easy载体。将CMV启动子和增强子控制的含GFP基因表达盒克隆入US2基因中，成功构建了 含GFP基因的转移质粒载体pGUS2GFP。用脂质体将其与CVI988株共转染CEF细胞，用96孔板稀释法得到纯 化的表达绿色荧光蛋白的重组CVI988病毒株rCVIGFP， 因的重组病毒在细胞上生长曲线与亲本毒CVI988类似， 体内分离到表达绿色荧光的病毒。 并分别测定其在体内和体外的生长情况。表达EGFP基 体外实验表明，1日龄腹腔接种该重组毒后，可以从鸡

The US2 gene nonessential for viral replication was amplified by polymerase chain reaction(PCR)with DNA from CEF cells infected by MDV vaccine strain CV1988 as template，and was cloned into T-easy vector；Th e expression cassette including GFP gene controlled by CM V promoter and enhancer was cloned into US2 gene to give rise to a tran sfer vector pGUS2GFP,then the complex of pGUS2GFP and DOTAP was transfected into CEF cells infected CVI988 strain virus．Recombinan t rCVIGFP expressing the green fluorescence protein was selected an d purified．Th e characterization of recombinant virus was evaluated n vivo and in vitro．Th e growth curve of recombinan t virus having EGFP gene are similar to that of the parent viru s in CEF cells．Th e recombinat virus expressing GFP could infect chicken and be reisolated from the chickens following intra-abdominal inoculation of rCVIGFP