H1N2亚型猪流感病毒NS1的原核表达及抗原性分析

Cloning,prokaryotic expression and antigenicity analyzing of NS1 Gene of H1N2 Swine influenza virus

为了进一步监控猪流感病毒对猪群的感染情况,研究对H1N2亚型猪流感病毒NS1基因进行RT-PCR扩增,并将其克隆到表达载体pET-30a(+)上构建重组质粒pET-NS1获得纯化产物,结果表明:NS1蛋白能够高效表达,与预期分子质量大小相符;经Western-blot分析,NS1蛋白能很好地与猪流感活病毒产生血清反应,而不能与灭活疫苗产生血清反应;被检血清稀释度为1∶640时,ELISA检测阳性血清的OD490值约为阴性血清的3倍,差异明显。说明NS1蛋白具有良好的血清学反应特异性。

To monitor swine flu virus infection between pigs,the NS1 gene of H1N2 subtype influenza A virus was amplified by RT-PCR,and then cloned into prokaryotic expression vector pET-30a（＋）.The recombinant plasmid pET-NS1 was obtained to produce purification of NS1 protein.The results showed that the recombinant protein of NS1 about 35 ku was produced.The Western-blot test showed that the recombinant protein reacted with serum produced from pigs immunized with the active virus,not with serum produced from pigs immunized with the inactivated vaccine.The OD490 value difference was distinctly that the positive sera 640 fold dilution is about 3 times of the negative sera.NS1 has good specificity of serological reactions,it is to establish swine flu antibody test kits of ELISA foundation.