猪呼吸道冠状病毒核衣壳蛋白基因的克隆及原核表达  

Cloning and Expression of Porcine Respiratory Coronavirus (PRCV) Nucleocapsid Protein

作　　者：曹敏杰[1] 郭川[1] 吴国平[1] 翁凌[1] 苏文金[1] 

机构地区：[1]集美大学生物工程学院,福建厦门361021

出　　处：《集美大学学报（自然科学版）》2005年第1期29-32,共4页Journal of Jimei University：Natural Science

基　　金：福建省科技攻关计划重点项目 (2003N083);厦门市科技攻关计划重点项目 (3502Z20031054);集美大学科研启动基金项目

摘　　要：以RT-PCR法从猪呼吸道冠状病毒(PRCV) 感染的ST细胞中得到核衣壳蛋白(N蛋白)cDNA, 并以此为模板利用PCR技术完成了N蛋白的基因扩增. 通过双酶切、连接、转化等过程构建了猪PRCVN蛋白基因的重组原核表达质粒, 表达产物经SDS PAGE、Westernblot检测被确认为GST融合的N蛋白. 表达蛋白与PRCV阳性血清间有良好的免疫结合性.Porcine respiratory coronavirus (PRCV) nucleocapsid protein (N protein) cDNA was (prepared) by RT-PCR using ST cells infected by PRCV.Using this cDNA as template,N protein full-length sequence DNA was amplified by PCR. Followed by restriction enzyme digestion,ligation and transformation,a recombinant pGEX 4T N protein plasmid was constructed.Expressed PRCV N protein occupied approximately 20 % of total bacteria protein and was mainly in soluble form as checked by SDS-PAGE. The protein was regarded as GST fused PRCV N protein after Western blot detection both using anti-GST monoclonal antibody and PRCV positive swine sera.

关 键 词：猪呼吸道冠状病毒 N蛋白 克隆 表达 

分 类 号：S852.65[农业科学—基础兽医学]