Baculovirus-encoded protein expression for epigenomic profiling in Drosophila cells

被引:4
作者
Bryson, Terri D. [1 ,2 ]
Weber, Christopher M. [1 ,3 ]
Henikoff, Steven [1 ,2 ]
机构
[1] Fred Hutchinson Canc Res Ctr, Div Basic Sci, Seattle, WA 98104 USA
[2] Howard Hughes Med Inst, Seattle, WA USA
[3] Univ Washington, Mol & Cellular Biol Program, Seattle, WA 98195 USA
基金
美国国家卫生研究院;
关键词
chromatin; epigenome; histone H2A. Z; cell lines; biotin-tagging; baculovirus; RECOMBINANT BACULOVIRUS; MAMMALIAN-CELLS; S2; CELLS; LINES; GENOME; MELANOGASTER; CHROMATIN; VECTORS; INSECT;
D O I
10.4161/fly.4.3.12177
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The expression and genome-wide mapping of epitope-tagged DN A-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.
引用
收藏
页码:258 / 265
页数:8
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