Literature DB >> 20495356

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

Terri D Bryson1, Christopher M Weber, Steven Henikoff.   

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.

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Year:  2010        PMID: 20495356     DOI: 10.4161/fly.4.3.12177

Source DB:  PubMed          Journal:  Fly (Austin)        ISSN: 1933-6934            Impact factor:   2.160


  4 in total

Review 1.  Generating and working with Drosophila cell cultures: Current challenges and opportunities.

Authors:  Arthur Luhur; Kristin M Klueg; Andrew C Zelhof
Journal:  Wiley Interdiscip Rev Dev Biol       Date:  2018-12-18       Impact factor: 5.814

2.  Cell-type-specific nuclei purification from whole animals for genome-wide expression and chromatin profiling.

Authors:  Florian A Steiner; Paul B Talbert; Sivakanthan Kasinathan; Roger B Deal; Steven Henikoff
Journal:  Genome Res       Date:  2012-01-04       Impact factor: 9.043

Review 3.  Cell lines.

Authors:  Lucy Cherbas; Lei Gong
Journal:  Methods       Date:  2014-01-13       Impact factor: 3.608

4.  H2A.Z nucleosomes enriched over active genes are homotypic.

Authors:  Christopher M Weber; Jorja G Henikoff; Steven Henikoff
Journal:  Nat Struct Mol Biol       Date:  2010-11-07       Impact factor: 15.369

  4 in total

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