公开(公告)号：US06342379B1

公开(公告)日：2002-01-29

申请号：US09459956

申请日：1999-12-13

申请人： Roger Y. Tsien ,  Jesus E. Gonzalez, III

发明人： Roger Y. Tsien ,  Jesus E. Gonzalez, III

IPC分类号： C12N1300

CPC分类号： G01N33/502 ,  C07D209/08 ,  C07D215/227 ,  C07D239/60 ,  C07D239/62 ,  C07D239/66 ,  C07D239/80 ,  C07D241/44 ,  C07D265/36 ,  C07D279/16 ,  C07D311/16 ,  C07D487/04 ,  C07F5/003 ,  C07F5/02 ,  C07F5/027 ,  C07F9/10 ,  C07F9/5728 ,  C07K2319/033 ,  C09B23/02 ,  G01N33/5005 ,  G01N33/5008 ,  G01N33/5044 ,  G01N33/5058 ,  G01N33/5061 ,  G01N33/5076 ,  G01N33/52 ,  G01N33/542 ,  G01N33/582 ,  G01N33/6872 ,  G01N33/92 ,  G01N2405/04 ,  G01N2500/10 ,  Y10S436/80 ,  Y10S436/805

摘要： Methods and compositions are provided for detecting changes in membrane potential in membranes biological systems. In one aspect, the method comprises; a) providing a living cell with a first reagent comprising a charged hydrophobic molecule which is typically a fluorescence resonance energy transfer (FRET) acceptor or donor, or is a quencher and is capable of redistributing within the membrane of a biological membrane in response to changes in the potential across the membrane; b) providing the cell with a second reagent that can label the first face or the second face of a biological membrane within the cell; c) detecting light emission from the first reagent or the second reagent. One aspect of this method involves monitoring membrane potential changes in subcellular organelle membranes in a living cells. Another aspect of the invention is the use of certain embodiments of the method for the screening of test chemicals for activity to modulate the activity of a target ion channel. Another aspect of the present invention is a transgenic organism comprising a first reagent that comprises a charged hydrophobic fluorescent molecule, and a second reagent comprising a bioluminescent or naturally fluorescent protein.

摘要翻译： 提供了用于检测膜生物系统中膜电位变化的方法和组合物。 一方面，所述方法包括：a）向活细胞提供包含带电疏水性分子的第一试剂，所述第一试剂通常为荧光共振能量转移（FRET）受体或供体，或者是猝灭剂，并且能够在膜内重新分布 的生物膜; b）向细胞提供可以标记细胞内的生物膜的第一面或第二面的第二试剂; c）检测来自第一个细胞膜的生物膜的光发射; 试剂或第二试剂。该方法的一个方面涉及监测活细胞中亚细胞细胞器膜中的膜电位变化。本发明的另一方面是使用该方法的某些实施方案来筛选测试化学品以用于调节 靶离子通道的活性。本发明的另一方面是转基因生物，其包含含有带电荷的氢的第一试剂 恐怖荧光分子和包含生物发光或天然荧光蛋白的第二试剂。