Lefevre Pascal, Bonifer Constanze
University of Leeds, St. James's University Hospital, Molecular Unit, Leeds, United Kingdom.
Methods Mol Biol. 2006;325:315-25. doi: 10.1385/1-59745-005-7:315.
Epigenetic processes involve alterations in the covalent modification of histones, which are driven by sequence-specific transcription factors recruiting the enzymatic machinery performing these reactions. Such histone modifications alter the charge or biochemical surface of the chromatin fiber and also serve as "docking" sites for transcription factor and chromatin remodelling complexes. Examining histone modification patterns is vital if we want to understand how transcription complexes establish patterns of gene expression. Histone modification can be quite localized and sometimes only involves a few nucleosomes. We therefore established a chromatin immunoprecipitation procedure that uses crosslinked chromatin allowing the isolation of small chromatin fragments while simultaneously minimizing histone movement during chromatin preparation.
表观遗传过程涉及组蛋白共价修饰的改变，这是由序列特异性转录因子招募执行这些反应的酶机制驱动的。这种组蛋白修饰改变了染色质纤维的电荷或生化表面，并且还作为转录因子和染色质重塑复合物的“停靠”位点。如果我们想了解转录复合物如何建立基因表达模式，那么研究组蛋白修饰模式至关重要。组蛋白修饰可能相当局部化，有时仅涉及少数核小体。因此，我们建立了一种染色质免疫沉淀方法，该方法使用交联染色质，允许分离小的染色质片段，同时在染色质制备过程中最大限度地减少组蛋白移动。
