DIAO Jun, ZHANG Xiu-Gen, CHEN Bu-Yan and CA Bao-Xiang. Construction of a Recom binant Herpesvirus of Turkey Expressing the Haem agglutinin-Neuram inidase Gene of Newcastle Disease Virus[J]. Virologica Sinica, 1999, 14(3): 259-264.
Citation: DIAO Jun, ZHANG Xiu-Gen, CHEN Bu-Yan, CA Bao-Xiang. Construction of a Recom binant Herpesvirus of Turkey Expressing the Haem agglutinin-Neuram inidase Gene of Newcastle Disease Virus .VIROLOGICA SINICA, 1999, 14(3) : 259-264.

表达新城疫病毒HN基因的重组火鸡疱疹病毒的构建

  • 应用聚合酶链反应技术,用设计带有限制性酶切位点的引物,特异地扩增从起始密码子开始的1.8kb新城疫病毒(NDV)血凝素神经氨酸酶(HN)基因开放式阅读框,然后插入pTK2B的NheⅠ位点,构建了含NDVHN基因的插入载体pTKHN1和pTKHN2,再与感染火鸡疱疹病毒(HVT)细胞的总DNA共转染鸡胚成纤维细胞(CEF),经有限稀释法和Dotblot筛选,得到含有HN基因的重组体rHVT1和rHVT2。经组织培养传代和Westernblot分析,表明重组体在感染细胞中表达了HN蛋白。重组体在CEF上的生长特性与亲本病毒相同,且在连续传代过程中保持稳定。为国内新城疫基因工程疫苗研制奠定了基础。

Construction of a Recom binant Herpesvirus of Turkey Expressing the Haem agglutinin-Neuram inidase Gene of Newcastle Disease Virus

  • A 1.8 kb fragment of Newcastle disease virus (NDV) haemagglutinin neuraminidase (HN) gene was amplified by polymerase chain reaction (PCR) with primers containing enzyme cleavage sites, and inserted into a nonessential gene of herpesvirus of turkey (HVT). Recombinant viruses were screened by Dot blot hybridization with digoxigenin labelled probe and purified by plaque formation technique. The expression of NDV HN gene can be detected by Western blotting in CEF cells infected with recombinant virus. Recombinant HVT strains were replicated stably in cell cultures.

  • 加载中
  • 加载中

Article Metrics

Article views(3886) PDF downloads(952) Cited by(0)

Related
Proportional views
    通讯作者: 陈斌, bchen63@163.com
    • 1. 

      沈阳化工大学材料科学与工程学院 沈阳 110142

    1. 本站搜索
    2. 百度学术搜索
    3. 万方数据库搜索
    4. CNKI搜索

    Construction of a Recom binant Herpesvirus of Turkey Expressing the Haem agglutinin-Neuram inidase Gene of Newcastle Disease Virus

    • 1. ColLege ofAnimal Medicine,Nanjing Agricultural University,Nanljlng 210095

    Abstract: A 1.8 kb fragment of Newcastle disease virus (NDV) haemagglutinin neuraminidase (HN) gene was amplified by polymerase chain reaction (PCR) with primers containing enzyme cleavage sites, and inserted into a nonessential gene of herpesvirus of turkey (HVT). Recombinant viruses were screened by Dot blot hybridization with digoxigenin labelled probe and purified by plaque formation technique. The expression of NDV HN gene can be detected by Western blotting in CEF cells infected with recombinant virus. Recombinant HVT strains were replicated stably in cell cultures.

    Relative (20)

    目录

    /

    DownLoad:  Full-Size Img  PowerPoint
    Return
    Return