Change in Linker DNA Conformation upon Histone HI.5 Binding to Nucleosome: Fluorescent Microscopy of Single Complexes

摘要

A method for fluorescently labeled DNA synthesis, which makes it possible to assemble mononu-cleosomes with 40 bp linkers, was developed. Cy3 and Cy5 labels were introduced into the linkers at distances of 10 bp before the first nucleotide and 15 bp after the last nucleotide of the nucleosome positioning DNA sequence, respectively. Without histone H1.5, fluorescence microscopy of single complexes revealed two equally probable states of nucleosomes. The states were different in the linker conformation