title: Purification of specific chromatin domains from single-copy gene loci in Saccharomyces cerevisiae.
abstract: Most methods currently available for the analysis of chromatin in vivo rely on a priori knowledge of putative chromatin components or their posttranslational modification state. The isolation of defined native chromosomal regions provides an attractive alternative to obtain a largely unbiased molecular description of chromatin. Here, we describe a strategy combining site-specific recombination at the chromosome with an efficient tandem affinity purification protocol to isolate a single-copy gene locus from the yeast Saccharomyces cerevisiae. The method allows robust enrichment of a targeted chromatin domain, making it amenable to compositional, structural, and biochemical analyses. This technique appears to be suitable to obtain a detailed description of chromatin composition and specific posttranslational histone modification state at virtually any genomic locus in yeast.
authors.affiliation: Biochemie-Zentrum Regensburg (BZR), Lehrstuhl Biochemie III, Universität Regensburg, Regensburg, Germany.
authors.forename: Stephan
authors.initials: S
authors.lastname: Hamperl
authors.forename: Christopher R
authors.initials: CR
authors.lastname: Brown
authors.forename: Jorge
authors.initials: J
authors.lastname: Perez-Fernandez
authors.forename: Katharina
authors.initials: K
authors.lastname: Huber
authors.forename: Manuel
authors.initials: M
authors.lastname: Wittner
authors.forename: Virginia
authors.initials: V
authors.lastname: Babl
authors.forename: Ulrike
authors.initials: U
authors.lastname: Stöckl
authors.forename: Hinrich
authors.initials: H
authors.lastname: Boeger
authors.forename: Herbert
authors.initials: H
authors.lastname: Tschochner
authors.forename: Philipp
authors.initials: P
authors.lastname: Milkereit
authors.forename: Joachim
authors.initials: J
authors.lastname: Griesenbeck
mesh_terms: D001671:Biochemistry / Q000379:methods*; D049109:Cell Proliferation; D002843:Chromatin / Q000378:metabolism*; D002846:Chromatography, Affinity; D004271:DNA, Fungal / Q000378:metabolism; D005656:Fungal Proteins / Q000378:metabolism; D018628:Gene Dosage; D005800:Genes, Fungal / Q000235:genetics*; D056426:Genetic Loci; D007074:Immunoglobulin G / Q000378:metabolism; D060328:Magnetic Phenomena; D008863:Microspheres; D012441:Saccharomyces cerevisiae / Q000166:cytology / Q000235:genetics* / Q000254:growth & development
chemical_list: D002843:Chromatin; D004271:DNA, Fungal; D005656:Fungal Proteins; D007074:Immunoglobulin G