Prokaryotic expression plasmid with nucleocapsid genes of TGEV/PEDV and co-expression of two genes
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摘要: 运用DNA重组技术将猪传染性胃肠炎病毒核衣壳基因、猪流行性腹泻病毒核衣壳基因编码最大局部亲水性抗原决定簇基因片段进行串联,克隆到原核表达载体pET-32a(+)中,构建预防仔猪冠状病毒性腹泻双价基因原核表达质粒。将该质粒转化至大肠杆菌BL21(DE3)中,经IPTG诱导后,SDS-PAGE电泳结果显示,双价重组基因表达出约86.8 kD的融合蛋白。Western-blotting检测结果表明,该融合蛋白具有良好的反应原性。Abstract: By using DNA recombination technology,nucleocapsid gene from swine transmissible gastroenteritis virus was lined with the gene fragment of the maximum local hydrophilic antigenic determinant of nucleocapsid gene from porcine epidemic diarrhea virus.The recombinant gene was cloned into the prokaryotic expression vector pET-32a(+).Then,the prokaryotic expression plasmid with the bivalent gene to prevent piglet’s coronaviral diarrhea was constructed.After transforming the plasmid into E.coil BL21(DE3),the bacteria were induced by IPTG with SDS-PAGE analysis showing the bivalent gene expressed efficiently,as well as,the western-blot test expressing the fusion protein.
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