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    Inducible fluorescence assay
    23.
    发明授权
    Inducible fluorescence assay 有权
    诱导荧光测定

    公开(公告)号：US07635598B2

    公开(公告)日：2009-12-22

    申请号：US11177998

    申请日：2005-07-08

    申请人： Ronald M. Cook ,  Eliana Saxon Armstrong ,  Hans E. Johansson

    发明人： Ronald M. Cook ,  Eliana Saxon Armstrong ,  Hans E. Johansson

    IPC分类号： G01N33/533 ,  G01N33/53 ,  C12Q1/26 ,  C12N1/21 ,  C12P21/00

    CPC分类号： C12Q1/26 ,  C09B11/08 ,  C12Q1/6818 ,  C12Q1/6897 ,  G01N33/542 ,  G01N2333/90209 ,  Y10S436/903 ,  Y10T436/13

    摘要： In one aspect, the invention provides fluorescent probes and assays. The probes include a fluorophore-quencher pair that undergoes a switch from dark to fluorescent in response to a reaction of the quencher. The switch of the probe from dark to fluorescent is typically mediated by an enzyme that acts directly or indirectly on the quencher, interfering with its ability to quench fluorescence emission from the fluorophore.In another aspect, the invention provides a reporter gene assay system and methods of using this system. The assay system includes a fluorophore-quencher probe and an enzyme that acts directly or indirectly on the quencher, increasing the fluorescent emission of the fluorophore. In still other aspects, the invention provides nucleic acid constructs and cells expressing the peptide products of these constructs.In assays of the invention, the presence of a target substance is detected by the switching of fluorescence mediated by the change in oxidation state of the quencher. The assay systems are of use in numerous assay formats, e.g., confirming the expression of the enzyme that acts on the quencher, and/or detecting a species conjugated to the enzyme.

    摘要翻译： 一方面，本发明提供了荧光探针和测定法。 探针包括响应于猝灭剂的反应经历从暗变为荧光的荧光团 - 猝灭剂对。 探针从黑暗到荧光的切换通常由直接或间接作用于猝灭剂的酶介导，干扰其从荧光团中淬灭荧光发射的能力。 另一方面，本发明提供报道基因测定系统和使用该系统的方法。 测定系统包括荧光团猝灭剂探针和直接或间接作用于猝灭剂的酶，增加荧光团的荧光发射。 在其它方面，本发明提供了表达这些构建体的肽产物的核酸构建体和细胞。 在本发明的测定中，通过由猝灭剂的氧化态的变化介导的荧光的转换来检测目标物质的存在。 测定系统可用于许多测定形式，例如确认作用于猝灭剂的酶的表达和/或检测与酶缀合的物质。