H7亚型禽流感病毒血凝素基因的真核表达载体的构建及鉴定  

Construction and Identification of Hemagglutinin Gene of Avian Influenza Virus Subtype H7 Eukaryotic Expression Vetor

作　　者：杨旭芹[1] 梁光军[1] 张小荣[1] 文其乙[1] 刘秀梵[1] 

机构地区：[1]扬州大学兽医学院畜禽传染病学农业部重点开放实验室,江苏扬州225009

出　　处：《中国家禽》2004年第z1期31-34,共4页China Poultry

摘　　要：参考已发表的H7亚型禽流感病毒(AIV)血凝素(HA)基因序列设计引物,以pUCH7为模板,经PCR扩增出一条1.7kb的HA全基因片段,将该片段定向克隆到真核表达质粒载体pcDNA3.1(一)中,转化大肠埃希氏菌DH5α,小量制备重组质粒pcDNA-HA,酶切鉴定正确后,转染COS-1细胞,经免疫荧光鉴定其体外表达情况。结果表明H7亚型AIV HA在COS-1细胞中获得了成功表达。用该重组质粒pcDNA-HA免疫BALB／C小鼠,制备免疫血清,经免疫印迹实验证实该H7亚型AIV HA在小鼠体内也得到了良好的表达。One 1.7 kb DNA fragment was amplified by PCR from pUCH7.The HA gene of H7 subtype AIV A/Afri. Star./Eng-Q/983/79(H7N1)was inserted into eukaryotic expression vector pcDNA3.l (-).The recombinant plasmid pcD-NA-HA was transfected into COS-1 cell by lipinfectin reagent after it was identified by restriction digestion.The result of IFA revealed the HA gene was expressed successfully in COS-1 cell.The BALB/C mice had been immunized by recombinant plasmid pcDNA-HA.And the result of Western Blot revealed the HA gene was also expressed successfully in mice and generated antibody against HA of H7 subtype AIV.

关 键 词：H7亚型禽流感病毒 血凝素基因 真核表达质粒载体 表达 

分 类 号：S83[农业科学—畜牧学]