Hershkovitz M, Riggs A D
Biology Department, Beckman Research Institute of the City of Hope, Duarte, California 91010, USA.
Methods. 1997 Feb;11(2):253-63. doi: 10.1006/meth.1996.0412.
Chromatin structure is becoming increasingly recognized as important for a full understanding of gene function and cell memory. With regard to cell memory, which involves the transfer of chromatin-encoded epigenetic information from one cell generation to another, the detailed structure of metaphase chromatin is of crucial importance. In this paper we describe methods for the use of dimethyl sulfate, DNase I, and potassium permanganate for in vivo footprinting and chromatin analysis, with special emphasis on studies of metaphase cells. We review the use of ligation-mediated PCR for the analysis of chromatin, including the human phosphoglycerate kinase promoter, and also report initial studies of a matrix attachment region near the human beta-interferon gene.
染色质结构对于全面理解基因功能和细胞记忆的重要性日益受到认可。关于涉及染色质编码的表观遗传信息从一个细胞世代传递到另一个细胞世代的细胞记忆,中期染色质的详细结构至关重要。在本文中,我们描述了使用硫酸二甲酯、DNase I和高锰酸钾进行体内足迹分析和染色质分析的方法,特别强调了对中期细胞的研究。我们回顾了连接介导的PCR在染色质分析中的应用,包括人类磷酸甘油酸激酶启动子,并且还报告了对人类β-干扰素基因附近一个基质附着区域的初步研究。