In vitro nucleosome assembly with plant histones

Abstract In vitro reconstitution of nucleosomes using cloned DNA sequences and animal histone proteins is straightforward and reproducible. However, there has been no report on any successful reconstitution experiment with plant histones. Here we report on the conditions under which reproducible in vitro reconstitution of different plasmids and purified plant histones takes place. Nucleosome formation is only observed with salt-extracted histones and in the presence of urea. In electron micrographs these reconstituted nucleosomes are similar to those assembled with animal histones. With the methods described prokaryotic as well as eukaryotic sequences cloned into plasmids can be packaged into nucleosomes of identical appearance. Southern transfer analyses revealed a repeat length of the reconstituted nucleosomes of 150 ± 8 bp. Nucleosome; DNA-histone interaction; Nucleosome reconstitution; Core histone; (Plant)