马立克氏病毒强毒GD0908株感染性克隆的构建

Construction of an Infectious Clone of a Virulent MDV Genome as Bacterial Artificial Chromosome

为研究马立克氏病毒(MDV)新型疫苗及MDV致病机理,将MDV强毒GD0908株的全基因组作为细菌人工染色体(BAC)转化进大肠杆菌,构建GD0908株的感染性克隆。利用同源重组将BAC载体插入MDV基因组的US2区,将包含BAC载体的MDV DNA电转化入大肠杆菌菌株DH10B,最后将鉴定成功包含GD0908株全基因组的BAC DNA转染鸡胚成纤维细胞(CEF),成功拯救出重组病毒,命名为rGD0908,其与父代病毒GD0908株在CEF细胞上的生长速度没有差异。该感染性克隆将为MDV的相关研究提供技术平台。

For better studying of the novel vaccine and pathogenesis about Marek＇ s disease virus, the complete genome of Marek＇ s disease virus （MDV） strain GD0908, which was isolated from Guangdong province in 2009, was cloned in Escherichia coli as a bacterial artificial chromosome （BAC）. BAC vector sequences were introduced into the US2 locus of the MDV genome by homologous recombination. The viral DNA containing the BAC vector was used to transform Escherichia coli strain of DH10B. Then the recombinant virus was successfully rescued by transfection of the recombinant BAC DNA into chicken embryo fibroblast （CEF）. This BAC viral clone was named rGD0908. There was no difference in growth ability in vitro between the BAC derived virus and its parental virus. With the powerful BAC manipulation system, the infectious clone will provide a platform for understanding the pathogenesis of the virulent MDV.