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Baculovirus-encoded protein expression for epigenomic profiling in Drosophila cells

机译:杆状病毒编码的蛋白表达用于果蝇细胞的表观基因组分析

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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.
机译:抗原决定簇标记的DN A和染色质结合蛋白在培养细胞中的表达和全基因组映射已成为表观基因组表征的有力策略,尤其是在果蝇中,果蝇现在可从许多组织中获得。然而,该策略依赖于建立转染的细胞系,这是费时的并且引入了可变性。在这里,我们显示了感染不同类型的果蝇细胞系后,可以高效生产杆状病毒编码的蛋白。使用染色质亲和纯化,我们显示在杆状病毒感染的细胞中产生的抗原决定簇标记的蛋白提供了组蛋白变体H2Av的全基因组谱,与质粒转染的细胞产生的谱图具有可比性。表达来自单个培养物的表观基因组分析的多个表位标记蛋白的能力,以及在各种果蝇细胞系中进行的能力,极大地扩展了表观基因组分析的范围。

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