摘要
利用GST融合基因表达系统表达GST-GP6融合蛋白,以质粒pMD19-T-ORF6为模板,扩增出ORF6基因片段,将其克隆至大肠杆菌表达载体pGEX-4T-2,将所构建的重组质粒pGEX-4T-2-ORF6转化E.coli ER2566,并诱导表达,采用GST蛋白纯化系统进行纯化,所得产物进行10%SDS-PAGE及Western blot鉴定。结果表明,大肠杆菌细胞经诱导高效表达出约42kD蛋白,其分子量与GST-GP6融合蛋白相符,表达产物以可溶的形式存在。Western blot分析表明,融合蛋白能够与斑点叉尾鮰病毒(Channel catfish virus,CCV)阳性血清发生特异性反应,表明该重组蛋白具有良好的抗原性和特异性。
Recombinant GST-GP6 fusion protein was expressed using GST gene fusion system.The gene ORF6 was amplified from plasmid pMD19-T-ORF6 and cloned into pGEX-4T-2 expression vector.The recombinant plasmid pGEX-4T-2-ORF6 was expressed in E.coli ER2566 cells and the products were purified by GST purifying system.The resolved GST-GP6 fusion protein on 10% SDS-PAGE showed a major band at position of 42kD and the fusion protein was recognized by antibody of channel catfish virus（CCV）.
出处
《湖北农业科学》
北大核心
2011年第5期1001-1003,1048,共4页
Hubei Agricultural Sciences
基金
国家自然科学基金项目(30700624)
