中文摘要：

根据GenBank中发表的禽流感病毒（AIV）血凝素（HA）序列设计了1对引物并分别引入NotⅠ和NcoⅠ酶切位点,用PCR方法扩增AIV HA基因片段。回收目的基因片段,并用NotⅠ/NcoⅠ限制性内切酶消化,经纯化后,将其定向克隆入果蝇表达载体pMT/B/V5中,构建重组表达载体pMT-HA。测序验证正确后,用脂质体转染法与辅助质粒pCoBlast共转染果蝇S2细胞,通过Blasticidin（杀稻瘟素）进行加压筛选,获得了抗性细胞S2-HA。提取S2-HA细胞的基因组DNA,PCR检测目的基因在果蝇S2细胞中的整合与表达。用终浓度500μmol/L的硫酸铜溶液诱导表达,收集无血清的细胞表达上清,样品浓缩后进行SDS-PAGE及Western Blotting检测。结果表明,成功构建了重组表达载体pMT-HA,转染细胞经10 mg/L的Blasticidin筛选及鉴定后获得了稳定表达HA的果蝇S2细胞株。

英文摘要：

Avian influenza（AI） is one of disease syndrome of birds which is caused by influenza A virus in avian（AIV）.The HA is the major surface antigen of influenza virus,which can induce the production of neutralizing antibodies to neutralize the infectivity of the virus.The study constructed a expressed vector（pMT-HA）.pMT-HA was transfected into S2 cells which was named S2-HA.S2-HA cell which could permanently express recombinant HA protein was established.The purity and speciality of HA were assessed using SDS-PAGE and Western blotting.