猪瘟病毒E2基因主要抗原区表达及Dot-ELISA建立研究  

Expression of The Major Antigen Region of E2 From CSFV and Preliminary Development of Dot-ELISA

作　　者：苏红辽 宗玉国 SU Hongliao;ZONG Yuguo(Huaxian Animal Hygienic Supervision Institute,Anyang 456400,China;Shandong Linqu County Comprehensive Administrative Law Enforcement Bureau,Weifang 262600,China)

机构地区：[1]河南省滑县动物卫生监督所,河南安阳456400 [2]山东省临朐县综合行政执法局,山东潍坊262600

出　　处：《中国猪业》2022年第4期64-68,共5页China Swine Industry

摘　　要：为建立基于猪瘟病毒E2基因主要抗原区表达产物的猪瘟抗体斑点杂交ELISA方法,应用RT-PCR方法扩增了猪瘟病毒E2基因的主要抗原区(dE2),经EcoR I、Xho I双酶切,与经同样双酶切的原核表达载体pET-28a (+)连接,加入适量IPTG诱导表达,SDS-PAGE和Western blotting分析结果表明,成功表达的重组蛋白分子量约32.4 KDa,能与CSFV阳性血清特异性结合,为CSFV抗体检测提供了有效工具。In order to acquire CSFV antigen and develop a Dot-ELISA for detecting antibody of CSFV,E2 of CSFV was amplified by RT-PCR,digested with EcoR I and Xho I and cloned into prokaryotic expression vector pET-28 a(+).Expression of E2 was induced by IPTG,and purified.The recombinant E2’ was analyzed by SDS-PAGE and Western blotting.The results showed that the expressed protein E2’ was about 32.4 KDa and reacted specifically with sera from pig infected with CSFV.Using E2 as coating antigen,a dot-ELISA was developed.It was an efficacious tool for detecting CSFV antibody.

关 键 词：猪瘟病毒 E2基因 原核表达 DOT-ELISA 

分 类 号：S828[农业科学—畜牧学] S852.651[农业科学—畜牧兽医]