中文摘要：

以马立克氏病病毒（MDV）CVI988株基因组为模板，利用PCR技术扩增出约2．7和3．0kb的基因片段，将上述片段同时插入pUC19中，获得约5．5kb MDV同源重组臂；以该基因片段的US2区的BglⅡ为插入位点，分别插入基因表达盒CMV-gpt-ployA和CMV—gfp-polyA，构建转移载体pUS-gpt-GFP，将该载体瞬时转染CHO细胞，在荧光显微镜下，可以观察到绿色荧光蛋白的表达；将该转移载体转染已用MDV CVI998株感染的次代CEF细胞，利用MX-HAT培养基筛选重组病毒，并用荧光显微镜挑选表达绿色荧光蛋白的蚀斑，结果获得重组病毒rMDV gptGFP。通过PCR检测和病毒生长测定，证明重组病毒获得纯化。

英文摘要：

Two fragments in the non-essential region of Marek＇s disease virus （MDV） were amplified by PCR from the genome of MDV CVI988 strain, and cloned into pUC 19 for generating the 5.5 kb homologous recombinant arms. A transfer vector pUS-gpt-GFP was constructed by inserting the expressing cassettes of the CMV-gpt-ployA or CMV-GFP-polyA, respectively, into the unique Bgl Ⅱ site of US2 region in the recombinant arms. The transfer vector was transiently transfected into CHO cells and the expression of the green fluorescence protein was observed under the fluorescence microscope. This transfer vector was then transfocted into CEF infected With MDV CVI988 strain. The recombinant CVI988 viruses expressing foreign genes, named rMDVgptGFP, were cloned by purifying the plagues expressing GFP in the MX-HAT selection medium. The purified recombinant viruses were further confirmed by PCR detection and growth of recombinant virus in selective and nonselective media.