公开(公告)号：US20050221402A1

公开(公告)日：2005-10-06

申请号：US10512497

申请日：2003-04-24

申请人： Eric Althoff ,  Virginia Cornish

发明人： Eric Althoff ,  Virginia Cornish

IPC分类号： C07K14/47 ,  C07K14/705 ,  G01N33/554 ,  C12N1/21 ,  C12N15/74 ,  G01N33/569

CPC分类号： C07K14/705 ,  C07K14/47 ,  C07K14/4705 ,  C07K2319/00

摘要： A transgenic bacterial cell comprising (a) a dimeric small molecule which comprises a first moiety known to bind a first receptor domain covalently linked to a second moiety known to bind a second receptor domain; (b) nucleotide sequences which upon transcription encode i) a first fusion protein comprising the first receptor domain, and ii) a second fusion protein comprising the second receptor domain; and (c) a reporter gene wherein expression of the reporter gene is conditioned on the proximity of the first fusion protein to the second fusion protein. The cell is also adapted for use in a method for identifying a molecule that binds to a known target in a bacterial cell from a pool of candidate molecules, and a method for identifying an unknown target receptor to which a molecule is capable of binding in a bacterial cell. Also described are compounds and kits for carrying out the methods.

摘要翻译： 一种转基因细菌细胞，其包含（a）二聚小分子，其包含已知结合与已知结合第二受体结构域的第二部分共价连接的第一受体结构域的第一部分; （b）转录后的核苷酸序列编码i）包含第一受体结构域的第一融合蛋白，和ii）包含第二受体结构域的第二融合蛋白; 和（c）其中报告基因的表达受到第一融合蛋白与第二融合蛋白接近度的调节基因的调节。 细胞还适用于鉴定从候选分子池中结合细菌细胞中的已知靶的分子的方法，以及用于鉴定分子能够在其中结合的未知靶受体的方法 细菌细胞。 还描述了用于实施方法的化合物和试剂盒。

公开(公告)号：US07575866B2

公开(公告)日：2009-08-18

申请号：US11219506

申请日：2005-09-01

申请人： Virginia Cornish ,  Michael Sheetz ,  Larry Miller

发明人： Virginia Cornish ,  Michael Sheetz ,  Larry Miller

IPC分类号： C12Q1/68 ,  C12Q1/26 ,  G01N33/53 ,  G01N33/573

CPC分类号： G01N33/582 ,  C12Q1/025 ,  C12Q1/32 ,  G01N33/5005 ,  G01N33/573 ,  G01N2333/906 ,  G01N2500/10

摘要： The present invention relates to methods and compositions which utilize binding relationships between detector molecules and their ligands for labeling molecules of interest in vivo. A “target” molecule is linked to a detector molecule (such as a protein or a nucleic acid) and the function and/or location of the target molecule is monitored by binding the detector molecule its ligand, which carries a detectable label. In particular embodiments of the invention, an intracellular fusion protein comprising a target protein and a detector protein is bound to a membrane-permeable, fluorescently-labeled ligand of the detector protein, thereby providing an adjunct or alternative to Green Fluorescent Protein.

摘要翻译： 本发明涉及使用检测器分子及其配体之间的结合关系的方法和组合物，用于在体内标记感兴趣的分子。 “靶”分子与检测器分子（例如蛋白质或核酸）连接，通过结合检测分子其配体，来监测靶分子的功能和/或位置，该配体携带可检测的标记。 在本发明的具体实施方案中，包含靶蛋白和检测蛋白的细胞内融合蛋白与检测蛋白的膜可渗透的荧光标记配体结合，从而提供绿色荧光蛋白的辅助物或替代物。

公开(公告)号：US20060211007A1

公开(公告)日：2006-09-21

申请号：US11219506

申请日：2005-09-01

申请人： Virginia Cornish ,  Michael Sheetz ,  Larry Miller

发明人： Virginia Cornish ,  Michael Sheetz ,  Larry Miller

IPC分类号： C12Q1/68 ,  G01N33/53 ,  C12Q1/26

CPC分类号： G01N33/582 ,  C12Q1/025 ,  C12Q1/32 ,  G01N33/5005 ,  G01N33/573 ,  G01N2333/906 ,  G01N2500/10

摘要： The present invention relates to methods and compositions which utilize binding relationships between detector molecules and their ligands for labeling molecules of interest in vivo. A “target” molecule is linked to a detector molecule (such as a protein or a nucleic acid) and the function and/or location of the target molecule is monitored by binding the detector molecule its ligand, which carries a detectable label. In particular embodiments of the invention, an intracellular fusion protein comprising a target protein and a detector protein is bound to a membrane-permeable, fluorescently-labeled ligand of the detector protein, thereby providing an adjunct or alternative to Green Fluorescent Protein.