Molecular beacons for detecting DNA binding proteins
Molecular beacons for detecting DNA binding proteins
复制标题
DOI：
10.1038/nbt0202-171
复制
发表时间：
2002-02-01
影响因子：
46.9
通讯作者：
Heyduk, E
中科院分区：
文献类型：
--
作者：
Heyduk, T;Heyduk, E
我们在这里报告了一种简单、快速、均质的荧光测定，即分子信标测定，用于检测和定量序列特异性 DNA 结合蛋白。该测定的核心特征是两个 DNA 片段的蛋白质依赖性关联，每个片段包含约一半的定义蛋白质结合位点的 DNA 序列。 DNA 片段的蛋白质依赖性关联可以通过任何基于邻近度的光谱信号进行检测，例如引入这些 DNA 分子的荧光染料之间的荧光共振能量转移 (FRET)。该测定是完全均质的，除了混合样品和测试溶液之外不需要任何操作。它在信号检测模式和荧光探针方面提供了灵活性，并且与多种蛋白质的多色同时检测兼容。该测定可用于研究和医学诊断以及针对 DNA 结合蛋白的药物的高通量筛选。
We report here a simple, rapid, homogeneous fluorescence assay, the molecular beacon assay, for the detection and quantification of sequence-specific DNA-binding proteins. The central feature of the assay is the protein-dependent association of two DNA fragments each containing about half of a DNA sequence defining a protein-binding site. Protein-dependent association of DNA fragments can be detected by any proximity-based spectroscopic signal, such as fluorescence resonance energy transfer (FRET) between fluorochromes introduced into these DNA molecules. The assay is fully homogeneous and requires no manipulations aside from mixing of the sample and the test solution. It offers flexibility with respect to the mode of signal detection and the fluorescence probe, and is compatible with multicolor simultaneous detection of several proteins. The assay can be used in research and medical diagnosis and for high-throughput screening of drugs targeted to DNA-binding proteins.