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Multiplex selection technique (MuST): an approach to clone transcription factor binding sites.

机译:多重选择技术(MuST):一种克隆转录因子结合位点的方法。

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摘要

We have used a multiplex selection approach to construct a library of DNA-protein interaction sites recognized by many of the DNA-binding proteins present in a cell type. An estimated minimum of two-thirds of the binding sites present in a library prepared from activated Jurkat T cells represent authentic transcription factor binding sites. We used the library for isolation of "optimal" binding site probes that facilitated cloning of a factor and to identify binding activities induced within 2 hr of activation of Jurkat cells. Since a large fraction of the oligonucleotides obtained appear to represent "optimal" binding sites for sequence-specific DNA-binding proteins, it is feasible to construct a catalog of consensus binding sites for DNA-binding proteins in a given cell type. Qualitative and quantitative comparisons of the catalogs of binding site sequences from various cell types could provide valuable insights into the process of differentiation acting at the level of transcriptional control.
机译:我们已经使用了多重选择方法来构建被细胞类型中存在的许多DNA结合蛋白识别的DNA-蛋白质相互作用位点的文库。由活化的Jurkat T细胞制备的文库中估计存在的结合位点的至少三分之二代表真实的转录因子结合位点。我们使用该文库来分离“最佳”结合位点探针,以促进因子的克隆并鉴定在激活Jurkat细胞后2小时内诱导的结合活性。由于获得的大部分寡核苷酸似乎代表了序列特异性DNA结合蛋白的“最佳”结合位点,因此在给定细胞类型中构建DNA结合蛋白的共有结合位点目录是可行的。来自各种细胞类型的结合位点序列目录的定性和定量比较可以提供有价值的见解,以了解在转录控制水平上起作用的分化过程。

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