鸡传染性腔上囊病病毒Gt株全基因组序列分析及真核表达载体的构建  

Analysis of the full length genome of infectious bursal disease virus strain Gt and construction of its eukaryotic expression vector

作　　者：祁小乐[1] 高玉龙[1] 高宏雷[1] 邓小芸[1] 张宁[1] 王晓燕[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：《中国兽医科学》2006年第10期820-826,共7页Chinese Veterinary Science

基　　金：国家重点基础研究发展计划(973)项目(2005CB523202)

摘　　要：采用一步法长距离RT-PCR克隆了IBDV Gt株全基因组,对其核苷酸和氨基酸序列进行了系统分析。在IBDV Gt株全基因组中引入分子标签,且在基因组两端分别引入锤头状核酶结构(HamRz)和丁肝病毒核酶结构(HdvRz)。将带有分子标签和核酶结构的IBDV基因组插入载体pCAGGS的β肌动蛋白启动子下游,构建了IBDV真核表达载体pCAGGmGtAHRT和pCAG-GmGtBHRT,为IBDV新型高效拯救平台的构建及基于此的病毒基因功能奠定了基础。The full length genome cDNA from strain Gt was cloned by the long accurate polymerase chain reaction（LA-PCR） in a single step amplification. The molecular biological information hidden in the full length genome was analysed in detail. EcoR Ⅴ site or Pst Ⅰ site, as the genetic tags, was introduced separately into the segment A or B of the IBDV Gt strain. The full length genome cDNA was flanked by hammerhead ribozyme（HamRz） and hepatitis D virus ribozyme（HdvRz） sequence. The full length genome eDNA containing genetic tags flanked by HamRz and HdvRz were arranged downstream of the β actin promoter of the vector pCAGGS. The constructed recombinant eukaryotic expression vector provided the basis for the development of infectious molecular clones of IBDV.

关 键 词：传染性腔上囊病病毒 Gt株 分子标签 核酶 真核表达载体 

分 类 号：S852.731[农业科学—基础兽医学] Q785[农业科学—兽医学]