传染性法氏囊病病毒超强毒株F9811VP_2基因的克隆及序列分析  

Cloning and Sequence Analysis of the VP_2 Gene of Very-Virulent Infectious Bursal Disease Virus F9811

作　　者：宋秀龙[1] 李春梅[2] 王笑梅[1] 陈冠春 陈奖励[1] 王秀荣[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,黑龙江哈尔滨150001 [2]延边大学农学院动物医学系,吉林龙井133400

出　　处：《中国兽医学报》2001年第3期223-226,共4页Chinese Journal of Veterinary Science

基　　金：欧盟合作项目! ( ERB 3 5 14 PL 972 776)

摘　　要：根据 NCBIG Gene Bank记载的传染性法氏囊病病毒超强毒 ( vv IBDV) OKYM株的核苷酸序列 ,设计并合成 1对特异性扩增 IBDV主要宿主保护性抗原 VP2 基因的引物。从 IBDV F981 1感染发病鸡法氏囊组织中提取病毒 RNA,经 RT-PCR扩增出约 1 .5kb的基因片段 ,采用平端连接法将此基因片段克隆至p UC1 1 9质粒的 Sma 位点上。经核苷酸序列测定 ,并与国内外其他 vv IBDV VP2 基因序列进行比较分析 ,发现 F981 1与 OKYM在 VP2 基因上有 1 8个核苷酸不同 ,但在氨基酸序列上仅 2 1 2位存在差异 ,从而从分子生物学角度证明 F981 1为 vv IBDV。对 1 4 3～ 3 82氨基酸区域所作系统进化树分析表明 ,F981 1、G92 0 1与OKYM、UK661、HK4 6相近 ,而 F950 2、G93 0 3与 OKYM、U K661、HK4 6较远 ,说明我国存在许多不同的vvThe major host protective antigen gene VP 2 of very virulent infectious bursal disease virus(vvIBDV) F9811 was amplified by RT PCR from bursa of one moribund chicken artificially infected vvIBDV F9811,with primers designed according to the published sequence of OKYM in NCBIG.The amplified fragment was about 1.5 kb,and cloned into the SmaⅠ site of pUC119 by blunt ligation.The VP 2 gene of F9811 was sequenced and compared with several vvIBDV strains abroad and domestic.F9811 was demonstrated to be vvIBDV by the fact that its VP 2 gene has 18 nucleotides,but only one amino acid at the 212 th position being different from OKYM.The phylogenetic tree based on the region 143 th^382 th nucleotide revealed that F9811 is a little nearer to OKYM,UK661,HK46 than F9502 and G9303.It implies that many different strains of vvIBDV may exist in China.

关 键 词：鸡 传染性法氏囊病病毒 超强毒株 基因克隆 序列分析 

分 类 号：S852.659.4[农业科学—基础兽医学] S188[农业科学—兽医学]