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中文摘要：
为探究鸡J亚群禽白血病病毒（ALV-J）的致病机理，采用PCR方法分4段分别扩增出J亚群禽白血病病毒安徽分离株AH-J11的前病毒cDNA，PCR产物经克隆鉴定，酶切后顺次连接，获得含有完整ALV-JAH-J11株前病毒cDNA的重组质粒，命名为pSKAH-J11。将AH-J11株全基因序列克隆至pBluescriptⅡSK（＋）载体中，得到重组质粒pBl-AH-J11，并将其转染鸡胚成纤维细胞（CEF）。通过间接免疫荧光试验，RT-PCR和Westernblot检测，并经测序验证比对拯救前后病毒的gp85基因序列，结果均表明拯救出了1株重组J亚群禽白血病病毒（rALV-J）。本研究成功构建了鸡ALV-J的感染性克隆并救获了其重组病毒，为研究禽白血病的致病机理提供了良好的反向遗传操作技术平台。
英文摘要：
In order to explore the pathogenesis of subgroup J avian leukosis virus（ALV-J）, a full- length infectious clone of ALV-J （pSK-AH-Jll） was constructed by combining of four fragments using PCR method from AH-Jll isolated from broiler in Anhui province, then the complete ge- nome was cloned into pBluescript II SK（＋） vector. The recombinant plasmid pB1-AH-J11 was transfected into chicken embryo fibroblast （CEF） cells and the recombinant ALV-J was rescued. The rescued ALV-J （rALV-J） was identified by RT-PCR, Western blot, indirect immunofluores- cence assay and sequence comparision of gp85 gene with AH-J11strain, respectively. The results showed that the recombinant rALV-J was rescued in CEF. This study provides a useful platform for investigation of the pathogenesis and molecular biology of ALV-J.
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