J亚群白血病病毒NX0101株感染性克隆化病毒的构建及其致病性

采用PCR方法分3段扩增出J亚群白血病病毒NX0 10 1株的前病毒cDNA ,PCR产物经克隆后顺次连接,获得一个含有完整ALV J前病毒cDNA的重组质粒,命名为pALV J NX。将此质粒DNA纯化后转染鸡胚成纤维细胞,以针对ALV J的单克隆抗体JE9对转染后的细胞作间接免疫荧光反应,证明获得了具有感染性的病毒。测定原始野毒和分子克隆化病毒的半数组织感染量(TCID50 ) ,分别人工接种1日龄商品代肉鸡并隔离饲养17周。接种野毒组死亡率为2 6 % ,髓细胞瘤发病率为2 4 %。接种分子克隆化病毒组死亡率为2 2 % ,髓细胞瘤发病率为2 2 %。结果表明,克隆化病毒具有天然病毒的致病性并对肉用型鸡表现致瘤性

Construction of infectious clone of subgroup J Avian leukosis virus strain NX0101 and its pathogenicity

By using PCR,3 fragments of provirus cDNA ofavian leukosis virus(ALV-J) strain NX0101 were amplified from the genomic DNA of ALV-J infected cells,and then combined in the right direction and sequences into recombinant plasmid pALV-J-NX, containing the whole genome of NX0101.After transfection of chicken embryo fibroblast (CEF) cells with plasmid pALV-J-NX DNA, the rescued virus was identified in CEF by indirect fluorescence antibody test with ALV-J specific monoclonal antibody JE9. The rescued virus could replicate in CEF at a titer of 10 5.6 /mL.The chicken experiment demonstrated that the rescued virus was still able to induce tumors in commercial meat-type broilers.