Mannose or xylose based positive selection
    1.
    发明授权
    Mannose or xylose based positive selection 失效
    甘露糖或木糖为阳性选择

    公开(公告)号:US5767378A

    公开(公告)日:1998-06-16

    申请号:US505302

    申请日:1995-10-03

    摘要: The present invention provides a method for identifying or selecting from a population of eukaryotic cells cultivated on or in a medium containing at least one compound, cells which have a metabolic advantage as a result of having being transformed, wherein: i) the cells are transformed with a nucleotide sequence or a co-introduced nucleotide sequence one of which comprises a region which: (a) encodes a protein which is involved in the metabolism of the compound, and/or (b) regulates the activity of the protein; and ii) the compound is mannose or xylose or a derivative or a precursor of these, or a substrate of the protein, or is capable of being metabolized by the transformed cells into such a substrate, with the proviso that the compound is not mannose when the protein is mannose 6 phosphate isomerase.The invention also includes a method according to the preceding paragraph wherein the compounds are not so limited with the proviso that an agent which reduces the toxicity to the cells of the compound is added to the medium. It is preferred that where a toxicity-reducing agent has been added to the culture medium, the compound is mannose and the nucleotide or co-introduced nucleotide sequence encodes mannose-6-phosphate isomerase.

    摘要翻译: PCT No.PCT / EP94 / 00575 Sec。 371 1995年10月3日第 102(e)日期1995年10月3日PCT 1994年2月28日PCT PCT。 公开号WO94 / 20627 1994年9月15日本发明提供了鉴定或选择在含有至少一种化合物的培养基上或培养基中培养的真核细胞群体的方法,所述培养基具有作为转化结果具有代谢优势的细胞,其中:i ),用核苷酸序列或共同引入的核苷酸序列转化细胞,其中之一包括以下区域:(a)编码参与化合物代谢的蛋白质,和/或(b)调节活性 蛋白质; 和ii)化合物是甘露糖或木糖,或其衍生物或前体,或蛋白质的底物,或能够被转化的细胞代谢成这样的底物,条件是该化合物不是甘露糖,当 蛋白质是甘露糖6磷酸异构酶。 本发明还包括根据前述段落的方法,其中化合物不限于此,条件是向培养基中加入降低化合物细胞毒性的试剂。 优选的是,当向培养基中加入毒性降低剂时,化合物是甘露糖,核苷酸或共同引入的核苷酸序列编码甘露糖-6-磷酸异构酶。