The Carnegie protein trap library: A versatile tool for Drosophila developmental studies
The Carnegie protein trap library: A versatile tool for Drosophila developmental studies
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DOI：
10.1534/genetics.106.065961
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发表时间：
2007-03-01
期刊：
影响因子：
3.3
通讯作者：
Spradling, Allan C.
中科院分区：
文献类型：
--
作者：
Buszczak, Michael;Paterno, Shelley;Spradling, Allan C.
后生动物的生理学依赖于错综复杂的基因表达模式，但这些模式仍然鲜为人知。利用果蝇转座子诱变，我们构建了7404个蛋白陷阱和增强子陷阱系的文库，卡内基文库，以便于在单细胞分辨率下进行基因表达图谱的绘制。通过对基因组插入位点进行测序，确定增强型绿色荧光蛋白(EGFP)外显子下游的剪接模式，以及分析卵巢和唾液腺的表达模式，我们发现600-900个不同的基因被困在我们的收集中。一个由244条线组成的核心集合捕获了不同的可识别蛋白质异构体，而在另外256个基因中发现了可能作为GFP增强子陷阱的插入。此外，还发现了至少8个新基因。我们的结果表明，卡内基收集将在细胞和发育生物学的不同领域作为一种有用的发现工具，并为通过插入蛋白质陷阱大幅增加果蝇蛋白质组的覆盖率提供了新的策略。
Metazoan physiology depends on intricate patterns of gene expression that remain poorly known. Using transposon mutagenesis in Drosophila, we constructed a library of 7404 protein trap and enhancer trap lines, the Carnegie collection, to facilitate gene expression mapping at single-cell resolution. By sequencing the genomic insertion sites, determining splicing patterns downstream of the enhanced green fluorescent protein (EGFP) exon, and analyzing expression patterns in the ovary and salivary gland, we found that 600-900 different genes are trapped in our collection. A core set of 244 lines trapped different identifiable protein isoforms, while insertions likely to act as GFP-enhancer traps were found in 256 additional genes. At least 8 novel genes were also identified. Our results demonstrate that the Carnegie collection will be useful as a discovery tool in diverse areas of cell and developmental biology and suggest new strategies for greatly increasing the coverage of the Drosophila proteome with protein trap insertions.