伪狂犬病病毒主要抗原表位基因KgE的表达及纯化  

Expression and Purification of Pseudorabies Virus Main Antigen Epitope Gene KgE

作　　者：周松峰[1,2] 吴健敏[2] 关忠宜[2] 廖文军[2] 白安斌[2] 

机构地区：[1]广西大学动物科学技术学院,广西南宁530005 [2]广西兽医研究所,广西南宁530001

出　　处：《动物医学进展》2011年第11期6-9,共4页Progress In Veterinary Medicine

基　　金：农业科技跨越计划项目(2009跨21);广西自然科学基金项目(2011GXNSFA018086)

摘　　要：从伪狂犬病病毒(PRV)基因组中克隆出KgE抗原表位基因,并将其插入到原核表达载体pET-Trx中,构建了原核表达质粒pET-Trx-KgE,将pET-Trx-KgE转化至BL21(DE3)plysS中,在IPTG诱导下进行表达。SDS-PAGE分析表明,KgE基因在BL21(DE3)plysS中获得高效表达,表达量占菌体总蛋白的32.8%,主要以包涵体形式存在,分子质量约为36ku。将表达的蛋白以亲和层析法进行纯化并通过谷胱甘肽还原法进行复性,纯化后蛋白纯度达到99.2%。Western blot检测发现,表达的KgE蛋白能够与PRV高免血清发生特异反应,但与猪繁殖与呼吸综合征病毒、猪瘟病毒等阳性血清不发生反应,表明KgE蛋白具有良好的抗原性。The main antigen epitope gene KgE was cloned from pig pseudorabies virus genome by conventional PCR method and inserted into the prokaryotic expression vector pET-trx,constructed recombinant plasmid pET-Trx-KgE and then transformed into competent cell BL21.The target protein was expressed with IPDG induction and obtained a high level expression by SDS-PAGE analysis,it took possession of whole bacterial proteins about 32.8% and presented as inclusion bodies mainly with weight 36 ku.The purity of expressed protein was reached 99.2% followed by chromatography purification and glutathione renaturation.There were no specific reactions of expressed KgE protein with positive sera of PRRSV or CSFV but with of PRV by Western blot analysis.It suggested that KgE protein has good antigenicity.

关 键 词：伪狂犬病病毒 KgE基因 原核表达 

分 类 号：S858.28[农业科学—临床兽医学] S852.659.1[农业科学—兽医学]