南阳师范学院学报2026，Vol.25Issue(1)：47-52,6.
猪伪狂犬病毒变异株gB基因主要抗原表位区的鉴定
Identification of Major Epitope Regions in the gB Gene of Variant Strains of Pseudorabies Virus in Pigs
摘要
Abstract
In this study,a segmented expression strategy was implemented to analyze the antigenic epitope regions of glycoprotein B(gB)in pseudorabies virus(PRV).Five pairs of specific primers were designed for the gB gene of PRV variant strains,and five gene fragments(gB-1 to gB-5)containing characteristic regions of different antigenic epitopes were successfully amplified.Each fragment was directly cloned into the pET-28a ex-pression vector to construct recombinant plasmids.After identification by double enzyme digestion and sequen-cing,the recombinant plasmids were transformed into BL21(DE3)competent cells,and the recombinant pro-teins were expressed through the IPTG induction system.SDS-PAGE analysis show that the gB-3(16 kDa),gB-4(30 kDa),and gB-5(30 kDa)fragments were highly expressed in Escherichia coli.The results of Western Blot analysis show that all three recombinant fusion proteins have specific reactions with monoclonal antibodies against His-tagged proteins.In addition,the proteins had specific immune reactions with PRV-positive pig sera collected from large-scale pig farms in different areas of Henan Province.Among them,The reactivity with the recombinant protein gB-5 is higher than that with gB-3 and gB-4,indicating that the gB-5 protein effectively retained the key antigenic epitopes of the natural gB protein of PRV.In addition,the gB-5 protein is located at positions 543～743 amino acid region of PRV gB protein and is relatively conserved among different PRV strains.In this study,the recombinant protein of the PRV gB gene with good antigenic activity was successfully screened out,providing an important basis and candidate antigen for the subsequent establishment of serological diagnostic methods for PRV.关键词
PRV变异株/gB基因/原核表达/抗原表位分析Key words
PRV variant strains/gB gene/prokaryotic expression/antigenic epitope analysis分类
农业科技引用本文复制引用
ZHAI Hongyue,WANG Ling,SONG Jiajing,WANG Jiabao,CAO Jiajia,LI Dandan,LENG Chaoliang..猪伪狂犬病毒变异株gB基因主要抗原表位区的鉴定[J].南阳师范学院学报,2026,25(1):47-52,6.基金项目
河南省科技攻关项目"基于M蛋白表位缺失的猪繁殖与呼吸综合征病毒标记疫苗株研制"(252102111005). （252102111005）
