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    • 1. 发明授权
    • Method for identifying products employing gene expression
    • 用于鉴定使用基因表达的产物的方法
    • US06576422B1
    • 2003-06-10
    • US09690391
    • 2000-10-17
    • Barry WeinsteinLorraine Holowach KellerSubba Reddy Palli
    • Barry WeinsteinLorraine Holowach KellerSubba Reddy Palli
    • C12Q168
    • C12Q1/6897
    • A method for identifying a product involves the steps of: (1) associating with the product a marker ligand; and (2) detecting the marker ligand in the product at a later point in time as a means of identifying the product by contacting the product with a detector composition. The detector composition comprises one or more first nucleotide sequences encoding one or more natural or synthetic ligand-dependent transcription factors, wherein said factors comprise at least one ligand binding domain, at least one DNA binding domain and at least one transactivation domain; and a second nucleotide sequence encoding a reporter gene under the regulatory control of a receptor response element or a modified or synthetic response element, and a second promoter. The method may also employ a corepressor or coactivator or a nucleotide sequence encoding the corepressor or activator Interaction between the marker ligand and ligand binding domain is highly specific and induces a change in the expression of the reporter gene, the change producing a detectable signal identifying the presence of the marker ligand in the product. The detector composition, a cell line containing the first and second nucleotide sequences, kits using them and products marked with specific marker ligands are useful in this method.
    • 用于鉴定产物的方法包括以下步骤:(1)与产物相关联的标记配体; 和(2)在稍后的时间点检测产品中的标记配体,作为通过使产物与检测器组合物接触来鉴定产物的手段。 检测器组合物包含编码一种或多种天然或合成配体依赖性转录因子的一种或多种第一核苷酸序列,其中所述因子包含至少一个配体结合结构域,至少一个DNA结合结构域和至少一个反式激活结构域; 以及在受体应答元件或修饰或合成应答元件的调节控制下编码报告基因的第二核苷酸序列和第二启动子。 该方法还可以使用共抑制因子或共激活物或编码共抑制因子或激活物的核苷酸序列。标记物配体和配体结合结构域之间的相互作用是高度特异性的并且引起报告基因表达的变化,该变化产生识别可检测信号 标记配体在产物中的存在。 检测器组合物,含有第一和第二核苷酸序列的细胞系,使用它们的试剂盒和标记有特异标记配体的产物在该方法中是有用的。