J亚群禽白血病病毒分离株HLJ09SH01株感染性克隆的构建及其致病性研究  

Construction of an infectious molecular clone of a subgroup J avian leukosis virus isolation HLJ09SH01 and its pathogenicity

作　　者：纪晓琳[1] 王琦[1] 高玉龙[1] 王永强[1] 秦立廷[1] 祁小乐[1] 高宏雷[1] 王笑梅[1] 

机构地区：[1]中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室/禽传染病研究室,黑龙江哈尔滨150001

出　　处：《中国预防兽医学报》2013年第1期15-18,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家自然科学基金(31072146);现代农业肉鸡产业技术体系建设(nycytx-42-G3-01);哈尔滨市科技攻关计划项目(2010AA6AN034)

摘　　要：为构建J亚群禽白血病病毒(ALV-J)分离株的感染性克隆并鉴定其对蛋鸡和肉鸡致病力的差异,本研究从黑龙江地区分离鉴定一株ALV-J分离株HLJ09SH01,以其核酸为模板,采用PCR方法分3段扩增其前病毒cDNA,PCR产物经克隆酶切后依次连接,获得一个含有完整ALV-J前病毒cDNA的重组质粒,命名为pBlue-HLJ09SH01。将其转染DF-1细胞,进行病毒拯救。通过间接免疫荧光、禽白血病抗原试剂盒检测及反转录酶活性试剂盒检验,表明拯救的病毒为ALV-J,命名为rHLJ09SH01。将其分别人工接种11日龄蛋鸡和肉鸡鸡胚,孵育出壳后隔离饲养32周。致瘤性试验结果表明,rHLJ09SH01可以导致57.9%的蛋鸡发生肿瘤,83.3%的肉鸡发生肿瘤。表明该感染性克隆具有亲本病毒的致病性。To construct the infectious clone of subgroup J avian leukosis virus （ALV-J） and identification of the oncogenicity of the rescued virus in layers and broilers, the sequence the provirus cDNA of ALV-J was amplified by PCR from ALV-J HLJ09SH01 isolated in Heilongjiang province and the infectious clone of pBlue-HLJ09SH01 was constructed based on the vector of pBluescript 1I KS（＋）. The rescued virus of rHLJ09SH01 was generated by transfecting pBlue-HLJ09SH01 into the DF-1 cells and identified by indirect immtmofluorescence assay, avian leukosis virus antigen test kit, reverse transcriptase assay. Furthermore, the 11 day old embryos of SPF layers and commercial broilers were inoculated with the rHLJ09SH01 via allantoic cavity and the results showed the rHLJ09SH01 caused typical tumors in SPF layer （57.9%） and broiler （83.3%） during 32 week period of observation after hatching, which demonstrated that the rescued virus possesses pathogenicity as parent virus.

关 键 词：J亚群禽白血病病毒 感染性分子克隆 致病性 

分 类 号：S852.65[农业科学—基础兽医学]