3′-U3区碱基突变不影响禽白血病病毒的体外复制能力  

The Mutations of 3′-U3 Region Make No Difference to ALV's Replication Capacity in vitro

作　　者：高雁怩 高奇[1] 李晓菲[1] 贠炳岭[1] 祁小乐[1] 王永强[1] 刘长军[1] 崔红玉[1] 张艳萍[1] 高宏雷[1] 王笑梅[1] 高玉龙[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免疫抑制病创新团队,哈尔滨150001

出　　处：《畜牧兽医学报》2015年第4期608-614,共7页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：国家自然科学基金(31372437);现代农业肉鸡产业技术体系(nycytx-42-G3-01)

摘　　要：为探索近年蛋鸡分离株中出现规律性突变的J群禽白血病病毒(ALV-J)3′-U3区对病毒体外复制能力的影响,以ALV-J原型毒株HPRS-103为骨架,利用融合PCR技术构建含蛋鸡分离株SD09DP04的3′-U3区的嵌合病毒的cDNA克隆。嵌合病毒的感染性克隆与原型毒株HPRS-103的感染性克隆分别在脂质体介导下转染DF-1细胞,对收获的病毒分别用间接免疫荧光试验、禽白血病抗原和反转录酶试剂盒检测,结果表明成功拯救到2株病毒。对拯救的2株病毒,进行其3′-U3区启动子活性和增强子活性的检测,以及复制动力学比较分析结果显示,出现特异性规律性突变的U3区对于原毒株的体外复制能力无显著影响。所构建的3′-U3区的嵌合病毒的成功拯救对于进一步探索近年ALV-J流行毒株其3′-U3区出现的规律性突变的功能及意义奠定了基础。In order to study the influence of the 3′-U3 region with regular mutations of epidemic J subgroup avian leukosis virus(ALV-J)strains isolated from egg-type flocks on its replication capacity in vitro,a full-length cDNA clone was constructed based on ALV-J prototype HPRS-103 strain infectious clone via replacing its 3′-U3 region by the corresponding part of egg-type isolate SD09DP04.Both the constructed chimeric infectious clone and the HPRS-103 infectious clone were transfected into DF1 cells by lipofectamine 2000,respectively.The two infectious clones were passaged to another generation.And the harvested viruses were respectively confirmed by indirect immunofluorescence(IFA),ALV-antigen test kit and reverse transcriptase activity test kit.Analyzing the 3′-U3region's promoter activity or enhancer activity and TCID50 growth curves showed that such U3 region with regular mutations had no evident effect on the virus' s replication capacity in vitro.The successful rescue of the chimeric virus plays the basic role in further exploration of the functions and the significance of the regular mutations in the 3′-U3 region in the epidemic ALV-J strains in recent years.

关 键 词：U3区 嵌合病毒 J亚群禽白血病病毒 生物学特性 

分 类 号：S852.65[农业科学—基础兽医学]