H17N10亚型蝙蝠流感病毒NS1蛋白的表达及其特性  

Expression and characterization of bat H17N10 influenza virus NS1 protein

作　　者：许凯迪 高俊娜[2] 侯天龙 刘芹防[2] 李泽君[2] 陈鸿军[2] 蒋文灿[1] 

机构地区：[1]四川农业大学动物医学院,四川成都611130 [2]中国农业科学院上海兽医研究所,上海200241

出　　处：《中国兽医科学》2016年第4期424-429,共6页Chinese Veterinary Science

基　　金：国家自然科学基金面上项目(31572502);中央公益性单位专项经费项目(2015JB15);中国农业科学院动物流感病原生态学创新团队专项经费

摘　　要：本研究通过构建H17N10亚型蝙蝠流感病毒NS1基因(BNS1)的原核表达载体。利用原核表达产物制备小鼠多抗血清,经间接免疫荧光试验检测其特异性。用真核表达载体p3Tag BNS1转染293T细胞进行免疫印迹检测。通过IFA及Western-blot检测,结果显示,BNS1原核表达产物为可溶性表达;BNS1多抗抗血清能与BNS1真核表达产物发生特异性结合;BNS1抗血清具备良好的免疫学活性。利用p RL-TK与p FL-IFNβ荧光素酶报告基因系统检测,结果表明,BNS1与H9N2 NS1均显著地降低IFN-β启动子活性。为下一步深入研究BNS1的生物学功能奠定了基础。NS1 gene of H17N10 bat influenza virus was constructed into the prokaryotic expression vector p Cold-TF.To prepare specific antisera to BNS1,the products were immunized into BALB/c mice at dose of 100 g per mouse.For identification of immunological characterization of BNS1, the gene was sub-cloned into p CMV-3Tag-2A vector and then transfected into 293 T cells.After 48 hours post-transfection,the protein were collected and identified by immunoblotting and by indirect immunofluorescence assay.The results showed that prokaryotic expression product was soluble and the antisera were specific and immunologic.Like H9N2 NS1,it showed that BNS1 significantly inhibited the activities of IFN-β promoter,which was detected by Dual-Luciferase Reporter Assay System.

关 键 词：蝙蝠流感病毒 H17N10亚型 NS1基因 原核表达 免疫学活性 

分 类 号：S852.659.5[农业科学—基础兽医学]