Chromatin structure mapping in Saccharomyces cerevisiae in vivo with DNase I.
Chromatin structure mapping in Saccharomyces cerevisiae in vivo with DNase I.
复制标题
使用 DNase I 绘制酿酒酵母体内染色质结构图。
DOI：
10.1093/nar/29.9.1943
复制
发表时间：
2001
影响因子：
14.9
通讯作者：
Simpson,RT
中科院分区：
文献类型：
--
作者：
Wang,X;Simpson,RT
Most methods for assessment of chromatin structure involve chemical or nuclease damage to DNA followed by analysis of distribution and susceptibility of cutting sites. The agents used generally do not permeate cells, making nuclear isolation mandatory.In vivomapping strategies might allow detection of labile constituents and/or structures that are lost when chromatin is swollen in isolated nuclei at low ionic strengths. DNase I has been the most widely used enzyme to detect chromatin sites where DNA is active in transcription, replication or recombination. We have introduced the bovine DNase I gene into yeast under control of a galactose-responsive promoter. Expression of the nuclease leads to DNA degradation and cell death. Shorter exposure to the active enzyme allows mapping of chromatin structure in whole cells without isolation of nuclei. The validity and efficacy of the strategy are demonstrated by footprinting a labile repressor bound to its operator. Investigation of the inter-nucleosome linker regions in several types of repressed domains has revealed different degrees of protection in cells, relative to isolated nuclei.
