VECTOR FOR IDENTIFICATION, SELECTION AND EXPRESSION OF RECOMBINANTS
    1.
    发明申请
    VECTOR FOR IDENTIFICATION, SELECTION AND EXPRESSION OF RECOMBINANTS 审中-公开
    鉴定,选择和表达重组体的载体

    公开(公告)号:US20110165583A1

    公开(公告)日:2011-07-07

    申请号:US13061640

    申请日:2009-09-02

    IPC分类号: C12Q1/68 C12N15/74 C12N15/63

    CPC分类号: C12N15/64 C12N15/10 C12N15/65

    摘要: A modified vector comprising a reporter gene having a STOP codon upstream of the multiple cloning site of the vector which is characterized in that the recombinant clones show fluoresce or show color in presence of inducer. A method for identification and selection of recombinant clones comprising the modified vector wherein the recombinant clones florescence or show color in a suitable suppressor strain of the STOP codon associated with the gene of interest. A method of preparation of recombinant clone comprising gene of interest and modified vector comprising amplification of gene of interest using specific primers containing STOP codon different from STOP codon used with reporter gene; cloning the amplified gene of interest in the modified vector; transformation of cloned modified vector in the STOP codon suppressor host cell wherein the STOP codon suppressor host cell is specific for STOP codon used with the gene of interest wherein the recombinant clones either fluorescence or show color depending upon the reporter gene used.

    摘要翻译: 包含在载体的多克隆位点上游具有一个末端密码子的报道基因的修饰载体,其特征在于重组克隆在诱导物存在下显示出荧光或显示出颜色。 用于鉴定和选择包含修饰载体的重组克隆的方法,其中所述重组克隆在与所关心的基因相关的STOP密码子的合适的抑制菌株中荧光或显示颜色。 包含目的基因和修饰载体的重组克隆的制备方法,其包括使用与报告基因使用的不同于STOP密码子的不同密码子的特异性引物扩增目标基因; 在经修饰的载体中克隆所扩增的目的基因; 克隆的修饰载体在STOP密码子抑制基因宿主细胞中的转化,其中所述的停止密码子抑制基因宿主细胞对于与感兴趣的基因一起使用的终止密码子是特异性的,其中所述重组克隆根据所使用的报道基因是荧光或显示颜色。