甲型H1N1流感病毒血凝素基因的原核表达及用于胶体金检测方法的研究  

Prokaryotic Expression of Hemagglutinin Gene of H1N1 Influenza A Virus and Its Preliminary Application in Colloidal Gold Immunochromatographic Assay

作　　者：王云龙[1,2] 郭文丽[1] 孙新城[3] 李玉林 李恒思 陈冬焕 王真 董彩文 刘旺根 

机构地区：[1]河南师范大学,河南新乡453007 [2]郑州职业技术学院,河南郑州450121 [3]郑州大学,河南郑州450001 [4]河南省生物工程技术研究中心,河南郑州450001

出　　处：《动物医学进展》2011年第7期1-4,共4页Progress In Veterinary Medicine

基　　金：河南省工程技术中心专项经费项目(102102313105);郑州市重大科技专项计划(093SGBS22027)

摘　　要：根据GenBank公布的甲型H1N1流感病毒血凝素(HA)基因序列,合成HA全长基因,构建原核表达载体pET-30 Xa/LIC-HA。阳性质粒转化E.coliBL21(DE3),IPTG诱导表达。对表达蛋白进行SDS-PAGE和Western blot检测的结果表明,HA基因获得表达,产物具有免疫学活性,分子质量约64 ku,以包涵体形式存在。以纯化的重组蛋白作为抗原初步建立了双抗原夹心检测甲型H1N1流感HA抗体的胶体金方法。该方法能检测出甲型H1N1流感阳性血清,为疫苗效果的评价奠定了基础。According to the relevant nucleotide sequence from GenBank,the hemagglutinin（HA） gene of H1N1 influenza A virus was synthetized.The product was used for constructing a prokaryotic recombinant plasmid pET-30 Xa/LIC-HA.The constructed vector was transformed to E.coli BL21（DE3） for expression under the induction of IPTG.The expressed product was identified by SDS-PAGE and Western blot.The target protein in the form of inclusion bodies,with a relative molecular mass of about 6 400,showed immunogenicity.A GICA for detecting antibodies against HA of H1N1 influenza A has been preliminarily developed with double-antigen sandwich assay using purified HA protein as antigen.Positive sera against H1N1 influenza A detected by GICA showed positive reaction.The method provided a solid basis for further research of the appraisal of vaccines.

关 键 词：甲型H1N1流感病毒 血凝素 原核表达 胶体金 

分 类 号：S858.285.3[农业科学—临床兽医学]