鸡传染性囊病病毒青岛分离株VP2基因的克隆及序列分析  

Cloning and sequence analysis of the VP2 gene of QingDao-IBDV isolate

作　　者：苏晓鸥[1] 乔俊文[1] 李玉荣[2] 赵德明[1] 杨利峰[1] 尹晓敏[1] 周向梅[1] 杨建民[1] 

机构地区：[1]中国农业大学动物医学院国家动物海绵状脑病实验室,北京海淀100193 [2]河北农业大学动物科技学院,河北保定071000

出　　处：《中国兽医杂志》2009年第7期6-10,共5页Chinese Journal of Veterinary Medicine

基　　金：国家科技支撑计划(2006BAD06A14);兽医微生物菌种资源标准化-朊病毒类资源标准化整理整合及共享(科技部;2005KDA21205-7)

摘　　要：为了克隆传染性囊病病毒青岛分离株(IBDV-QD)的VP2基因及对其高变区序列进行分析,首先根据GenBank上已经发表的传染性囊病病毒(IBDV)UK661株的核苷酸序列,设计并合成了一对特异性的扩增IBDV-QDVP2基因的引物。以IBDV-QD株基因组为模版,利用RT-PCR技术扩增出长约1.5kb的基因片段,将其连接到pGEM-T-Easy载体上。经酶切分析、PCR鉴定及核苷酸序列测定,成功获得该分离株VP2基因的重组质粒。将IBDV-QD株VP2基因的高变区核苷酸序列及推导出的氨基酸序列与GenBank中公布的14株IBDV相应序列应用软件进行同源性比较,并构建系统进化树。同源性比较和进化树分析表明,IBDV-QD与IBDV超强毒株(vvIBDV)关系最近,与标准强毒株、弱毒株、变异株相差较远;并且我国不同地区的vvIBDV存在差异,这为我国传染性囊病的流行病学调查奠定了基础。To clone the VP2 gene from infectious bursal disease virus of QingDao isolated strain （IB- DV-QD） and analyze the sequence of hypervariable region of VP2, a pair of specific primers used to ampli- fy complete eDNA of VP2 gene of IBDV-QD was firstly designed and synthesized according to the pub- lished sequence of IBDV UK661 strain on GenBank. Using^the genome of IBDV-QD as template, approxi- mate 1.5 kb DNA fragment was obtained by RT-PCR technique, and then VP2 eDNA was connected with pGEM-T-Easy vector. Positive clones were successfully identified by PCR and EcoR I enzyme digestion and then sequenced. The obtained nucleotide sequence and deduced amino acid sequence were compared with those of 14 IBDV strains published in GenBank , and phytogenetic tree based on the amino acid se~ quences of hypervariable region of VP2 was built with software. The comparable results demonstrated that IBDV-QD strain had a higher similarity to very virulent infectious bursal disease virus（vvIBDV）,lower to standard, weak virulent and variant strains. The results also show that there are different vvIBDV strains in different areas of our country, which lays the foundation in epidemioligical investigation into infectious bursal disease epidemiology.

关 键 词：IBDV-QD分离株 鸡传染性囊病病毒 VP2基因 序列分析 

分 类 号：S852.65[农业科学—基础兽医学]