鸭瘟病毒gB基因B细胞和T细胞表位的预测及其主要抗原域基因的原核表达  

Prediction of B-and T-cell epitopes of gB gene of duck plague virus and prokaryotic expression of its major antigen determinants

作　　者：林丹[1] 汪铭书[1,2] 程安春[1,2] 朱德康[1,2] 贾仁勇[2] 罗启慧[2] 刘菲[2] 陈孝跃[1,2] 

机构地区：[1]四川农业大学动物医学院禽病防治研究中心,四川雅安625014 [2]动物疫病与人类健康四川省重点实验室,四川雅安625014

出　　处：《中国兽医科学》2009年第10期866-872,共7页Chinese Veterinary Science

基　　金：教育部"长江学者和创新团队发展计划"创新团队项目(IRT0848);现代农业产业技术体系建设专项项目(nycytx-45-42);教育部高等学校科技创新工程重大项目培育资金项目(706050)

摘　　要：应用生物信息学工具对本实验室鉴定的鸭瘟病毒(DPV)CHv毒株gB基因(GenBank登录号:EF608147)编码蛋白进行了B、T细胞表位预测,对其主要抗原域基因进行了PCR扩增,构建了原核表达载体pET-28a-gBM,并在大肠杆菌BL21(DE3)中诱导表达。结果表明,重组菌可表达分子质量约46 ku的重组融合蛋白,表达蛋白以包涵体形式存在,约占菌体总蛋白的30%。Western-blot分析显示,表达蛋白能够被兔抗DPV多克隆血清特异识别,证实该氨基酸片段具有较强的免疫原性。B- and T-ceil epitopes of duck plague virus（DPV） gB gene（GenBank accession No. EF608147） were predicted by bioinformatics softwares. A fragment encoding gB major epitopes was ampli-fied by PCR and subcloned into the prokaryotic expression vector pET-28a（＋）. The constructed recombi- nant was transformed into Escherichia coli BL21（DE3） competent cells and induced to express. The results showed that the recombinant protein with molecular weight of 46 ku was expressed in inclusion bodies and accounted for 30% of the total proteins. Western-blot analysis showed that the expressed protein could be recognized specifically by polyclonal rabbit anti-DPV serum,indicating that the expressed protein possessed strong reactmogenlcity.

关 键 词：鸭瘟病毒 GB基因 抗原表位 原核表达 

分 类 号：S852.659.1[农业科学—基础兽医学]