两株不同亚型禽流感病毒NS1基因的表达及抗原性检测  

Expression and Detection of Nonstructural Protein NS1 Gene of Two Avian Influenza Virus Strains

作　　者：孙进华[1] 于志丹[1] 曲哲会[1] 张雅为[1] 王君伟[1] 

机构地区：[1]东北农业大学动物医学院,哈尔滨150030

出　　处：《中国动物检疫》2006年第9期28-30,共3页China Animal Health Inspection

摘　　要：为进一步研究禽流感病毒非结构蛋白NS1的功能及其在临床中的应用,本试验分别对H5N1、H9N2两株不同亚型禽流感病毒的NS1基因进行了原核表达及抗原性检测。实验分别用限制性内切酶EcoRI和XhoI消化不同亚型NS1基因的重组质粒pMD-18T-NS1(H9N2)和pMD-18T-NS1(H5N1),获得目的片段NS1,然后将NS1基因克隆到原核表达载体pProEXHTc中。将重组质粒pProc-NS1(H9N2、H5N1)转化DH5α感受态细胞,用1mmol/LIPTG诱导表达,表达产物进行SDS-PAGE检测,目的蛋白分子量大小为30ku,与预期结果相符。West-ern-blot检测表明所表达的蛋白能与NS1的多克隆抗血清反应,并且存在交叉反应。In order to research the function and application of nonstructural （NS1） of the avian influenza, the nonstructural protein NS1 gene of two strains of the avian influenza virus（H5N1 .H9N2） was expressed and detected. The recombinant plasimid of pMD-18T-NS1 （H9N2） and pMD-18T-NS1 （H5N1） was digested by EcoRI or XhoI respectively. The products were cloned onto the expression vector pProEXHTc respectively. Recombinant plasmid pPro-NS1 （H9N2、H5N1） was transformed into E.coli DH5αcompetent cells respectively and induced with 1mmol/L IPTG.The target protein was produced.SDS-PAGE test indicated that the molecular weight of the expressed protein was 30ku as expected.Western-blot test indicated that the expressed protein can react with the NS1 polyclonal antibody of the influenza virus, and can cross reaction each other.

关 键 词：禽流感病毒 非结构蛋白 基因表达 检测 

分 类 号：S855.3[农业科学—临床兽医学] TS201.21[农业科学—兽医学]