作　　者： ; ; ; ; ; ; ;

机构地区： 华南农业大学动物科学学院

出　　处： 《华南农业大学学报》 2006年第3期93-96,共4页

摘　　要： 采用RT-PCR方法扩增了H9N2亚型禽流感病毒(AIV)广东株的血凝素基因HA.将HA基因克隆到笔者所设计的原核表达载体pMBX上,成功构建重组表达质粒pMBX-HA.将其转化到E.coliBL-21感受态细胞中表达,经SDS-PAGE可检测到相对分子质量约为96 200的融合蛋白,表达产物占菌体总蛋白的29.5%.经蛋白可溶性分析看出,融合蛋白90%以上是以可溶形式表达的.W estern-b lot证实,可溶表达的融合蛋白与H9N2 AIV阳性血清具有良好的免疫反应性.将可溶表达的融合蛋白用体积分数为50%的N i-NTA树脂过柱纯化,用融合蛋白作抗原,通过ELISA方法,能特异性地检测出禽流感阳性血清. The HA gene of H9N2 subtype avian influenza virus （A1T） isolated from Guangdong Province was successfully amplified by RT-PCR. The HA fragment was inserted into plasmid pMBX to obtain recombinant plasmid in which HA gene was fused into. The pMBX-HA was transformed into E. coli BL-21 （DE3） to induce HA gene fusion expression with 1 mmol/L IPTG. The fusion protein band with relative molecular mass of 96 200 was detected on the SDS-PAGE gel, and the soluble expession products accounted for 29.5 % of the total E. coli proteins. The soluble form of the expression products was up to 90% the fusion protein. The expression products specifically reacted with the antisera against H9N2 subtype avian influenza virus. The soluble expression products was purified by 50% （φ） Ni-NTA affinity chromatography and used as an antigen, AIV antibody could be detected by ELISA method.

关 键 词： 禽流感病毒 基因 高效可溶表达 免疫原性

领　　域： [农业科学] [农业科学] [农业科学]