Analysis of chromatin structure in vivo

Methods : a Companion to Methods in Enzymology
J S MymrykT K Archer

Abstract

A number of important nuclear processes including replication, recombination, repair, and transcription involve the interaction of soluble nuclear proteins with DNA assembled as chromatin. Recent progress in a number of experimental systems has focused attention on the influence chromatin structure may exert on gene regulation in eukaryotes. With the advent of new technologies for the analysis of chromatin structure in vivo, studies evaluating the influence of chromatin structure on gene transcription have become feasible for a number of systems. This article serves as an introduction to the use of restriction endonucleases to define nucleosomal organization and characterize changes in this organization that accompany transcriptional activation in vivo. The procedure includes the isolation of intact transcriptionally competent nuclei, limited digestion with specific restriction endonucleases, and purification of the DNA. This DNA serves as the substrate for a linear amplification using single primers that generate enzyme-specific DNA fragments, which are then resolved by electrophoresis. Specific examples related to our studies of the influence of chromatin structure on steroid hormone regulation of transcription from the mouse...Continue Reading

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Citations

Sep 15, 1999·Proceedings of the National Academy of Sciences of the United States of America·S R Cherry, D Baltimore
Nov 6, 2001·Molecular Biology of the Cell·B J Deroo, T K Archer
Mar 20, 2012·Biochimica Et Biophysica Acta·Heather A KingTrevor K Archer

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