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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
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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1，2
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（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；2.黑龙江省农业科学院 博士后科研工作站，黑龙江 哈尔滨150001）
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摘要：
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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pCAGGmGtBHRT，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。 |
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关键词：
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传染性腔上囊病病毒；Gt株；分子标签；核酶；真核表达载体 |
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中图分类号：S 852.731：Q 785 文献标识码：
A 文章编号：1673-4696(2006)10-0820-07 |
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Analysis of the full length genome of infectious bursal disease virus
strain Gt and construction of its eukaryotic expression vector |
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QI Xiao-le1，GAO Yu-long1，GAO
Hong-lei1，DENG Xiao-yun1，
ZHANG Ning1，WANG Xiao-yan1，WANG Xiao-mei1，2 |
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(1.Division of Avian Infectious Diseases,National Key Laboratory of Veterinary Biotechnology/Harbin Veterinary
Research Institute,Chinese Academy of Agricultural Sciences,Harbin 150001, China; 2.Postdoctor Scientific
Research Workstation, Heilongjiang Academy of Agricultural Sciences, Harbin 150001, China) |
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Abstract: 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. EcoRⅤ site or PstⅠsite, 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 cDNA 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. |
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Key words: infectious bursal disease virus (IBDV); Gt strain; genetic tags; ribozyme; eukaryotic expression vector |
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