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目的 克隆甲型流感病毒血凝素基因HA1区及构建其真核表达载体。方法 从接种人流感病毒株A/PR/ 8/ 34(H1N1)的鸡胚尿囊液中提取病毒RNA ,用特异引物进行RT PCR ,扩增血凝素基因HA1区。将所扩增片断克隆入真核表达质粒载体pcDNA3.1(+) ,转化感受态大肠杆菌JM 10 9并筛选阳性克隆。结果 经双酶切、PCR及测序鉴定证实血凝素基因HA1区的真核表达载体构建成功。结论 甲型流感病毒血凝素的HA1区是与宿主细胞膜表面受体结合的部位并起着诱导机体产生保护性抗体的作用 ,HA1区的克隆和其真核表达质粒的构建将为防治病毒侵入宿主细胞及核酸疫苗的研究打下基础。
Objective To clone the HA1 region of influenza A virus hemagglutinin gene and construct its eukaryotic expression vector. Methods The virus RNA was extracted from chick embryo allantoic fluid inoculated with human influenza virus strain A / PR / 8/34 (H1N1). The specific primers were used for RT PCR to amplify the hemagglutinin gene HA1. The amplified fragment was cloned into the eukaryotic expression plasmid vector pcDNA3.1 (+), transformed into competent E. coli JM109 and screened positive clones. Results The eukaryotic expression vector of hemagglutinin HA1 region was successfully constructed by double enzyme digestion, PCR and sequencing. Conclusion The HA1 region of influenza A virus hemagglutinin binds to the receptor on the surface of host cell membrane and plays a role in inducing the body to produce protective antibody. The cloning of HA1 region and construction of its eukaryotic expression plasmid will be helpful for preventing virus invasion Host cells and nucleic acid vaccine laid the foundation for the study.