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Fly (Austin). 2010 Jul-Sep;4(3):258-65. doi: 10.4161/fly.4.3.12177. Epub 2010 Jul 1.

Baculovirus-encoded protein expression for epigenomic profiling in Drosophila cells.

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  • 1Basic Sciences Division, Fred Hutchinson Cancer Research Center, Seattle, WA, USA.

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.

PMID:
20495356
[PubMed - indexed for MEDLINE]

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