In vitro DNA-binding profile of transcription factors: methods and new insights.

In vitro DNA-binding profile of transcription factors: methods and new insights.
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转录因子的体外 DNA 结合谱:方法和新见解。

DOI:
10.1530/joe-11-0010
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发表时间:
2011-07
期刊:
The Journal of endocrinology
影响因子:
--
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--
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其他
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转录因子 (TF) 的 DNA 结合特异性对细胞生理学、细胞发育和进化具有广泛的影响。然而,大多数已知转录因子的 DNA 结合特异性仍然未知。 TF 蛋白的特异性由其与所有可能的结合位点的相对亲和力决定。近年来,几种体外技术的发展允许高通量测定 TF 对所有可能的 k bp 长 DNA 序列的相对结合亲和力,从而极大地促进了许多已知 TF 的 DNA 结合特异性的表征。所有可被具有各种结合亲和力的 TF 结合的 DNA 序列形成其 DNA 结合谱 (DBP)。 DBP对于生成准确的DNA结合模型、识别全基因组中所有DNA结合位点和转录因子的靶基因以及构建转录调控网络具有重要意义。本研究回顾了这些技术,特别是两项主要技术:双链 DNA 微阵列和通过指数富集结合平行 DNA 测序技术 (SELEX-seq) 的配体系统进化。
The DNA-binding specificity of transcription factors (TFs) has broad impacts on cell physiology, cell development and in evolution. However, the DNA-binding specificity of most known TFs still remains unknown. The specificity of a TF protein is determined by its relative affinity to all possible binding sites. In recent years, the development of several in vitro techniques permits high-throughput determination of relative binding affinity of a TF to all possible k bp-long DNA sequences, thus greatly promoting the characterization of DNA-binding specificity of many known TFs. All DNA sequences that can be bound by a TF with various binding affinities form their DNA-binding profile (DBP). The DBP is important to generate an accurate DNA-binding model, identify all DNA-binding sites and target genes of TFs in the whole genome, and build transcription regulatory network. This study reviewed these techniques, especially two master techniques: double-stranded DNA microarray and systematic evolution of ligands by exponential enrichment in combination with parallel DNA sequencing techniques (SELEX-seq).
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