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    Method for selecting high-expressing host cells
    5.
    发明授权
    Method for selecting high-expressing host cells 失效
    选择高表达宿主细胞的方法

    公开(公告)号：US5561053A

    公开(公告)日：1996-10-01

    申请号：US286740

    申请日：1994-08-05

    申请人： Craig W. Crowley

    发明人： Craig W. Crowley

    IPC分类号： C12N15/09 ,  C07K14/715 ,  C07K16/42 ,  C12N5/00 ,  C12N5/10 ,  C12N9/64 ,  C12N9/72 ,  C12N15/64 ,  C12N15/67 ,  C12N15/85 ,  C12P21/02 ,  C12R1/91 ,  C12P21/00 ,  C07H21/04 ,  C12N15/11

    CPC分类号： C12N9/6456 ,  C07K14/7151 ,  C07K16/4283 ,  C12N15/64 ,  C12N15/67 ,  C12N15/85 ,  C07K2319/00 ,  C12N2830/42 ,  C12N2840/20 ,  C12N2840/44

    摘要： A method for selecting recombinant host cells expressing high levels of a desired protein is described. This method utilizes eukaryotic host cells harboring a DNA construct comprising a selectable gene (preferably an amplifiable gene) and a product gene provided 3' to the selectable gene. The selectable gene is positioned within an intron defined by a splice donor site and a splice acceptor site and the selectable gene and product gene are under the transcriptional control of a single transcriptional regulatory region. The splice donor site is generally an efficient splice donor site and thereby regulates expression of the product gene using the transcriptional regulatory region. The transfected cells are cultured so as to express the gene encoding the product in a selective medium comprising an amplifying agent for sufficient time to allow amplification to occur, whereupon either the desired product is recovered or cells having multiple copies of the product gene are identified.

    摘要翻译： 描述了选择表达高水平所需蛋白质的重组宿主细胞的方法。 该方法利用携带包含可选择基因（优选可扩增基因）的DNA构建体的真核宿主细胞和向可选基因提供3'的产物基因。 可选择的基因位于由剪接供体位点和剪接受体位点限定的内含子内，并且可选择的基因和产物基因处于单个转录调节区的转录控制之下。 剪接供体位点通常是有效的剪接供体位点，从而使用转录调节区调节产物基因的表达。 培养转染的细胞以在包含扩增剂的选择性培养基中表达编码产物的基因足够长的时间以允许发生扩增，从而鉴定所需产物或识别具有多个拷贝的产物基因的细胞。