传染性喉气管炎病毒WG株糖蛋白gD基因主要抗原区的表达  

Expression of major antigenic region of glycoprotein gD gene of infectious laryngotracheitis virus WG strain

作　　者：魏秀英[1,2] 石星明[2] 张晶[2] 王玫[2] 赵妍[2] 胡顺磊[2] 崔红玉[2] 全炎铭[2] 闫帅[2] 陈洪岩[1,2] 王云峰[1,2] 

机构地区：[1]东北农业大学生命科学学院,黑龙江哈尔滨150030 [2]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》2011年第2期188-192,共5页Chinese Veterinary Science

基　　金：国家高技术研究发展计划(863)项目(2003AA213020)

摘　　要：为表达传染性喉气管炎病毒(ILTV)gD蛋白,根据GenBank中登录的ILTV全基因组序列(EU675324)设计了1对引物,进行gD全基因的PCR扩增,产物经纯化后连接至pGEM-T Easy载体,进行序列测定,采用DNAStar软件分析gD蛋白的抗原性,选择抗原性强的第256～405aa的编码片段设计引物并进行PCR扩增,结果获得截短的gD基因,将其克隆至原核表达载体pET-32a中,并转化E.coli BL21(DE3)。经IPTG诱导后,获得大小为43.5ku的重组蛋白,命名为r-gD。Western-blot分析结果表明,r-gD具有较强的抗原性。To express the gD protein of infectious laryngotracheitis infectious virus（ILTV）,a pair of primers was designed to amply gD gene according to the ILTV genome sequence（EU675324） available in GenBank.The PCR amplified gD gene was purified and ligated to pGEM-T Easy vector for sequencing.According to sequencing results and DNAStar software analysis of the antigenicity of gD protein,the fragment encoding 256 aa to 405 aa which is more antigenic was selected for design primers.The fragment of gD gene was cloned into pET-32a vector and transformed into Escherichia coli BL21（DE3）.After induction with IPTG,a recombinant protein with the size of 43.5 ku was expressed and named r-gD.Western-blot analysis showed that the r-gD had strong antigenicity,which provided a foundation for preparation of monoclonal antibodies and the development of diagnostic methods for ILTV.

关 键 词：传染性喉气管炎病毒 糖蛋白D 序列分析 蛋白质免疫印迹 

分 类 号：S852.659.1[农业科学—基础兽医学]