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从SELEX实验推断蛋白质-DNA相互作用参数。

Inferring protein-DNA interaction parameters from SELEX experiments.

作者信息

Djordjevic Marko

机构信息

Faculty of Biology, Department of Chemistry and Physics, University of Belgrade, Arkansas State University, Studentski trg 16, 11000 Belgrade, Serbia.

出版信息

Methods Mol Biol. 2010;674:195-211. doi: 10.1007/978-1-60761-854-6_12.

Abstract

Systematic Evolution of Ligands by EXponential enrichment (SELEX) is an experimental procedure that allows extraction, from an initially random pool of oligonucleotides, of the oligomers with a high binding affinity for a given molecular target. The highest affinity binding sequences isolated through SELEX can have numerous research, diagnostic, and therapeutic applications. Recently, important new modifications of the SELEX protocol have been proposed. In particular, a suitably modified SELEX experiment, together with an appropriate computational procedure, allows inference of protein-DNA interaction parameters with up to now unprecedented accuracy. Such inference is possible even when there is no a priori information on transcription factor binding specificity, which allows accurate predictions of binding sites for any transcription factor of interest. In this chapter we discuss how to accurately determine protein-DNA interaction parameters from SELEX experiments. The chapter addresses experimental and computational procedure needed to generate and analyze appropriate data.

摘要

指数富集的配体系统进化技术(SELEX)是一种实验方法,它能够从最初随机的寡核苷酸库中筛选出对特定分子靶标具有高结合亲和力的寡聚物。通过SELEX分离出的具有最高亲和力的结合序列可用于众多研究、诊断和治疗应用。最近,人们提出了对SELEX方案的重要新改进。特别是,经过适当修改的SELEX实验,结合适当的计算程序,能够以前所未有的精度推断蛋白质-DNA相互作用参数。即使在没有关于转录因子结合特异性的先验信息的情况下,这种推断也是可能的,这使得能够准确预测任何感兴趣的转录因子的结合位点。在本章中,我们将讨论如何从SELEX实验中准确确定蛋白质-DNA相互作用参数。本章阐述了生成和分析适当数据所需的实验和计算程序。

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