【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.更多还原

【Abstract】 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.更多还原