马疱疹病毒1型gB糖蛋白线性B细胞表位的预测与鉴定  

Prediction and Identification of Linear B-cell Epitopes in gB Glycoprotein of Equine Herpesvirus Type 1

作　　者：李静[1] 刘建华[1] 付强[1] 宋焕堂 范斌[1] 胡月[1] 冉多良[1] 

机构地区：[1]新疆农业大学动物医学学院,乌鲁木齐830052

出　　处：《中国畜牧兽医》2017年第5期1484-1490,共7页China Animal Husbandry & Veterinary Medicine

基　　金：国家科技支撑计划项目(2012BAD46B04-03)

摘　　要：试验旨在应用生物信息学技术综合分析马疱疹病毒1型(equine herpesvirus 1,EHV-1)gB糖蛋白,预测B细胞表位,筛选出具有潜在诊断价值的线性B细胞表位。将EHV-1gB糖蛋白的基因序列输入DNAStar软件中的Protean工作区中,经参数综合比较分析筛选潜在的B细胞表位,克隆、表达预测表位的基因片段,利用表达的融合蛋白作为抗原与马疱疹病毒阳性血清反应。经预测分析,gB糖蛋白的B细胞表位可能位于第6—10、23—32、53—65、72—98、111—120、152—166和173—180位氨基酸区域。本试验成功构建并原核表达含7个潜在B细胞表位的融合蛋白。Western blotting试验结果显示,其中5个融合蛋白能被马疱疹病毒阳性血清识别。本试验利用生物信息学技术结合分子生物学技术成功筛选到5个潜在的B细胞表位,为EHV-1表位诊断、表位疫苗抗原的设计奠定了技术基础。The study was aimed to predict B cell epitopes in gB glycoprotein of equine herpesvirus type 1（EHV-1）with bioinformatics,and select epitopes which had potential diagnostic value.The DNA fragments of gB glycoprotein were predicted by protean of DNAStar software.Screening potential B cell epitopes after parameter comparison,the target B cell epitopes were selected,cloned and expressed.The expressed fusion proteins serviced as an antigen were used to react with equine herpesvirus positive serum to screen and identify antigenic epitopes.The results showed that according to predictive and analysis,the areas of amino acid from 6to 10,23 to 32,53 to 65,72 to 98,111 to 120,152 to 166,173 to 180might be gB glycoprotein B cell epitopes.Seven epitopes were successfully cloned into a prokaryotic expression vector,and confirmed by DNA sequencing.After expression and purification,Western blotting was performed to detect the antigen,which could be recognized by equine herpesvirus positive sera.Bioinformatics technology and molecular biology techniques were used to successfully screen five potential B cell epitopes,which provided the foundation for the diagnosis of EHV-1and design of the epitope vaccine.

关 键 词：B细胞表位 抗原表位预测 原核表达 鉴定 

分 类 号：S852.652[农业科学—基础兽医学]