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    • 1. 发明授权
    • Transcription factor-DNA binding assay
    • 转录因子-DNA结合测定
    • US5563036A
    • 1996-10-08
    • US235503
    • 1994-04-29
    • Michael G. PetersonVijay R. BaichwalBerta Strulovici
    • Michael G. PetersonVijay R. BaichwalBerta Strulovici
    • G01N33/566C12N15/09C12Q1/68C12Q1/70G01N33/50
    • G01N33/5008C12Q1/6811C12Q1/6813C12Q1/6876C12Q1/701C12Q2600/136
    • Pharmacological agents useful in the diagnosis or treatment of disease associated with the expression of a gene are identified in high throughput drug screening assays. The methods involve combining a labeled transcription factor, a nucleic acid coupled to a ligand, a candidate pharmacological agent and a receptor immobilized on a solid substrate, such as a microtiter plate, filter, or bead. The nucleic acid has at least that portion of a nucleotide sequence naturally involved in the regulation of the transcription of the gene which is necessary for sequence-specific interaction with the transcription factor. The resultant combination is incubated under conditions whereby the receptor is bound to the ligand and, but for the presence of said candidate pharmacological agent, the transcription factor is sequence-specifically bound to the nucleic acid. Unbound transcription factor is then removed or washed from the solid substrate and labelled, sequence-specifically bound transcription factor is detected. Incubates which include candidate agents which alter transcription factor binding deviate from control incubates in terms of label signal--typically, binding is disrupted and the signal is diminished. In a preferred embodiment, the entire process is performed by a computer-controllable electromechanical robot with an axial rotatable arm.
    • 用于诊断或治疗与基因表达相关的疾病的药理学试剂在高通量药物筛选试验中鉴定。 所述方法包括将标记的转录因子,与配体偶联的核酸,候选药理学试剂和固定在固体基质如微量滴定板,过滤器或珠上的受体组合。 该核酸至少具有与转录因子的序列特异性相互作用所必需的天然参与调节基因转录的核苷酸序列的那部分。 将所得组合在受体结合于配体的条件下孵育,并且对于所述候选药理学试剂的存在,转录因子与核酸序列特异性结合。 然后从固体底物中除去或洗涤未结合的转录因子,并检测到标记的序列特异性结合的转录因子。 包括候选药物的孵化物,其通过改变转录因子结合偏离对照培养物的标签信号(通常结合)被破坏并且信号减弱。 在优选实施例中,整个过程由具有轴向可旋转臂的计算机可控机电机器人执行。