【关键词】 非洲猪瘟; VP72; 原核表达; 基因免疫; ELISA;

【英文关键词】 African swine fever virus; VP72; Prokaryotic expression; ELISA;

【中文摘要】 非洲猪瘟(African Swine Fever, ASF)是由ASFV引起的猪的急性、烈性传染病,以全身出血、呼吸障碍和神经症状为主要特征,强毒株感染的发病率和死亡率可达100%,对养猪业危害巨大,是各国严密防范的传染病,也是世界猪病研究的重点。目前我国尚无本病的发生。但随着国际贸易和人员交往的日益频繁,该病传入我国的风险是存在的,因此建立快速诊断方法十分必要。 为了建立ASFV特异抗体检测方法,本研究根据国外发表的ASFV VP72基因序列设计引物,以英国动物健康研究所提供的含VP72基因的质粒为模板,扩增VP72基因及其可溶性抗原片断VP72S,将其插入pUC载体中进行测序,所获序列与已发表的ASFV-VP72(Ba71株)完全相同。将测序正确的VP72和VP72S基因克隆入pGEX-6p-1载体中GST基因的下游,重组大肠杆菌用IPTG诱导表达,并采用不同方法纯化得GST融合蛋白。将VP72基因克隆入真核表达pcDNA3中,重组载体用多聚乙酸亚胺包裹后基因免疫8周龄小鼠,用制备的血清对原核表达产物进行Western-blotting验证,结果表明,本实验原核表达的重组VP72是正确的...

【英文摘要】 African swine fever (ASF) is a highly lethal viral hemorrhagic disease in domestic pigs. The morbidity and mortality of the disease caused by virulent virus can be 100%, with characteristic of multiple hemorrhages, breathing difficulty and nerve symptoms. It presents a great potential harm to swine industry worldwide. China is presently free of the disease, but the risk of its entrance is possible due to increasing international trade. Thus, it is necessary to develop quick and accurate diagnostic meth...