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使用连接介导的PCR方法进行体内基因组足迹分析。

In vivo genomic footprinting using LM-PCR methods.

作者信息

Tagoh Hiromi, Cockerill Peter N, Bonifer Constanze

机构信息

Molecular Medicine Unit, St. James's University Hospital, University of Leeds, United Kingdom.

出版信息

Methods Mol Biol. 2006;325:285-314. doi: 10.1385/1-59745-005-7:285.

Abstract

Epigenetic regulatory proteins such as transcription factors, chromatin components, and chromatin modification activities alter gene activity during development. The means by which alterations in these factors influence gene expression is poorly understood, but information of this kind is essential if we want to reprogram the epigenotype of specific cell types in a directed fashion. To facilitate chromatin structure-function analysis, we have developed a relatively simple procedure that uses magnetic beads to perform ligation-mediated polymerase chain reaction in solid phase. In this chapter, we describe detailed procedures for the examination of chromatin fine-structure and nucleosome positioning as well as changes in transcription factor binding-site occupancy during cellular differentiation.

摘要

表观遗传调控蛋白,如转录因子、染色质成分和染色质修饰活性,在发育过程中改变基因活性。这些因子的改变影响基因表达的方式尚不清楚,但如果我们想要定向重编程特定细胞类型的表观基因型,这类信息至关重要。为便于染色质结构-功能分析,我们开发了一种相对简单的程序,该程序使用磁珠在固相进行连接介导的聚合酶链反应。在本章中,我们描述了用于检查染色质精细结构和核小体定位以及细胞分化过程中转录因子结合位点占用情况变化的详细程序。

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