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Analysis of the expression patterns, Subcellular localisations and interaction partners of drosophila proteins using a pigp protein trap library

机译:使用Pigp蛋白陷阱库分析果蝇蛋白的表达模式,亚细胞定位和相互作用伴侣

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摘要

Although we now have a wealth of information on the transcription patterns of all the genes in the Drosophila genome, much less is known about the properties of the encoded proteins. To provide information on the expression patterns and subcellular localisations of many proteins in parallel, we have performed a large-scale protein trap screen using a hybrid piggyBac vector carrying an artificial exon encoding yellow fluorescent protein (YFP) and protein affinity tags. From screening 41 million embryos, we recovered 616 verified independent YFP-positive lines representing protein traps in 374 genes, two-thirds of which had not been tagged in previousPelement protein trap screens. Over 20 different research groups then characterized the expression patterns of the tagged proteins in a variety of tissues and at several developmental stages. In parallel, we purified many of the tagged proteins from embryos using the affinity tags and identified co-purifying proteins by mass spectrometry. The fly stocks are publicly available through the Kyoto Drosophila Genetics Resource Center. All our data are available via an open access database (Flannotator), which provides comprehensive information on the expression patterns, subcellular localisations and in vivo interaction partners of the trapped proteins.Our resource substantially increases the number of available protein traps in Drosophila and identifies new markers for cellular organelles and structures.
机译:尽管我们现在掌握了果蝇基因组中所有基因转录模式的大量信息,但对编码蛋白质的特性了解甚少。为了提供有关许多蛋白质并行表达模式和亚细胞定位的信息,我们使用携带带有编码黄色荧光蛋白(YFP)和蛋白质亲和标签的人工外显子的杂种ggyBac载体,进行了大规模蛋白质陷阱筛选。通过筛选4100万个胚胎,我们回收了616个经过验证的独立YFP阳性品系,这些品系代表374个基因中的蛋白质陷阱,其中三分之二在以前的Pelement蛋白质陷阱筛选中未标记。然后,超过20个不同的研究小组表征了标记蛋白在各种组织中以及几个发育阶段的表达模式。同时,我们使用亲和标签从胚胎中纯化了许多标签蛋白,并通过质谱鉴定了共纯化蛋白。蝇果可以通过京都果蝇遗传资源中心公开获得。我们所有的数据都可以通过开放访问数据库(Flannotator)获得,该数据库提供了有关被困蛋白质的表达模式,亚细胞定位和体内相互作用伙伴的全面信息。我们的资源大大增加了果蝇中可用的蛋白质陷阱数量并确定了新的细胞器和结构的标记。

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