H7亚型禽流感病毒HA1基因在重组杆状病毒中的表达及其表达产物的抗原性  

Expression of HA1 gene of avian influenza virus H7 subtype in recombinant baculovirus and analysis of antigenicity of the expressed HA1 protein

作　　者：贾珊珊[1] 孙凌霜[1] 齐岩[1] 孙进华[1] 姜亦飞[1] 王君伟[1] 

机构地区：[1]东北农业大学动物医学院,黑龙江哈尔滨150030

出　　处：《中国兽医科学》2007年第3期232-235,共4页Chinese Veterinary Science

基　　金：国家科技攻关计划项目(2004BA519A23)

摘　　要：从pMD18-T-HA阳性质粒中扩增出H7亚型禽流感病毒(AIV)HA1基因,并亚克隆至昆虫杆状病毒转移载体pBlueBacHis2A,将筛选的重组质粒命名为pBlueBacHisH7-HA1,与线性化杆状病毒DNA(Bac-N-BlueDNA)共转染sf9昆虫细胞,挑取蓝色蚀斑,经3次蚀斑纯化,得到重组杆状病毒rpBlueHisH7HA1。Western-blotting和间接免疫荧光试验结果显示,HA1在昆虫细胞中得到表达,且表达产物具有抗原性。抗原特异性试验证明,重组HA1蛋白与鸡H5、H9亚型AIV抗血清不发生交叉反应。血凝试验证明,重组HA1蛋白具有良好的血凝性。The HA1 gene of avian influenza virus H7 subtype was amplified from recombinant plasmid pMD-18-T-HA by PCR. The PCR product was subcloned into pBlueBacHis2A vector. The recombinant transfer vector pBlueHisH7-HA1 was screened at random. After pBlueHisH7-HA1 was co-transfected with Bac-N-Blue DNA by cationic liposome-mediated transfection, and was purified by three cycles of plaque assay with chromogenic substracte X-gal in low temperature-melting agarose, a recombinant baculovirus, designated as rpBlueHisH7-HAl,was obtained. Western-blotting and indirect immunofluorescence tests showed the HA1 gene was well expressed,and the expressed product had antigenicity. An antigen specific test showed that the recombinant did not have any cross-reaction with chicken H5 and H9 antisera. A hemagglutinin test certified that it had good hemagglutination.

关 键 词：禽流感病毒 HA1基因 重组杆状病毒 H7亚型 

分 类 号：S852.659.5[农业科学—基础兽医学] Q786[农业科学—兽医学]