【作者】 张纪元； 崔治中； 丁家波； 姜世金；

【Author】 ZHANG Ji - yuan,CUI Zhi - zhong~*,DING Jia - bo,JIANG Shi -jing (College ofAnimal Science andTechnology,ShandongAgricultural University,Tai’an271018,China)

【机构】 山东农业大学动物科技学院；

【摘要】 采用PCR方法分3段扩增出J亚群白血病病毒NX0101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26%,髓细胞瘤发病率为24%。接种分子克隆化病毒组死亡率为22%,髓细胞瘤发病率为22%。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。更多还原

【Abstract】 By using PCR,3 fragments of provirus cDNA ofavian leukosis virus(ALV -J) strain NX0101 were amplified from the genomic DNA ofALV - 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 viruswas identified in CEF by indirect fluorescence antibody testwith ALV - J specific monoclonal antibody JE9.The rescued virus could replicate in CEF at a titerof 105.6/mL.The chicken experiment demonstrated that the rescued virus was still able to induce tumors in commercial meat - type broilers.更多还原