title: Baculovirus-encoded protein expression for epigenomic profiling in Drosophila cells.
abstract: The expression and genome-wide mapping of epitope-tagged DNA- and chromatin-binding proteins in cultured cells has become a powerful strategy for epigenome characterization, especially in Drosophila, where cell lines derived from numerous tissues are now available. However this strategy relies on establishing transfected cell lines, which is time-consuming and introduces variability. Here we show that baculovirus-encoded proteins can be efficiently produced following infection of Drosophila cell lines of different types. Using chromatin affinity purification, we show that epitope-tagged proteins produced in baculovirus-infected cells provide genome-wide profiles of the histone variant H2Av that are comparable to those produced by plasmid-transfected cells. The ability to express multiple epitope-tagged proteins for epigenome analysis from a single culture, and to do this in a variety of Drosophila cell lines, significantly extends the range of epigenome analysis.
authors.affiliation: Basic Sciences Division, Fred Hutchinson Cancer Research Center, Seattle, WA, USA.
authors.forename: Terri D
authors.initials: TD
authors.lastname: Bryson
authors.forename: Christopher M
authors.initials: CM
authors.lastname: Weber
authors.forename: Steven
authors.initials: S
authors.lastname: Henikoff
mesh_terms: D000818:Animals; D016367:Baculoviridae / Q000502:physiology*; D019921:Biotinylation; D002460:Cell Line; D004330:Drosophila / Q000235:genetics / Q000378:metabolism*; D057890:Epigenomics / Q000379:methods*; D006657:Histones / Q000378:metabolism; D010957:Plasmids; D014176:Protein Biosynthesis
chemical_list: D006657:Histones