斑点叉尾鮰病毒ORF6基因的高效可溶表达及抗原性分析

摘要

Recombinant GST-GP6 fusion protein was expressed using GST gene fusion system. The gene ORF6 was amplified from plasmid pMDI9-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).%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.