Reconstitution of chromatin complexes from high-performance liquid chromatography-purified histones.
We describe a method to reconstitute chromatin complexes from reversed-phase high-performance liquid chromatography (HPLC)-purified histones. The complexes reconstituted in this way exhibit the same structural characteristics as their equivalent native counterparts. Furthermore, this method works independently of the acid- or salt-extracted origin of the histones used for the HPLC fractionation. The potential of this method for the reconstitution of chromatin particles consisting of sequence-defined DNA templates and well-defined histone variants and/or their posttranslationally modified isoforms is discussed.
Ausió
J
J
Department of Biochemistry and Microbiology, University of Victoria, British Columbia, Canada. jausió@uvic.ca
Moore
S C
SC
D001671:Biochemistry / Q000379:methods*; D002843:Chromatin / Q000737:chemistry*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004247:DNA / Q000737:chemistry* / Q000378:metabolism; D018983:DNA Footprinting / Q000379:methods; D015246:Deoxyribonuclease EcoRI / Q000737:chemistry / Q000378:metabolism; D003850:Deoxyribonuclease I; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D006657:Histones / Q000302:isolation & purification; D009707:Nucleosomes / Q000737:chemistry; D014461:Ultracentrifugation / Q000379:methods
D002843:Chromatin; D006657:Histones; D009707:Nucleosomes; D004247:DNA; D015246:Deoxyribonuclease EcoRI; D003850:Deoxyribonuclease I