Profiling patterned transcripts in Drosophila embryos
Profiling patterned transcripts in Drosophila embryos
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DOI：
10.1101/gr.84402
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发表时间：
2002-07-01
期刊：
影响因子：
7
通讯作者：
Letsou, A
中科院分区：
文献类型：
--
作者：
Simin, K;Scuderi, A;Letsou, A
在这里，我们描述了一种高通量筛选，以分离早期胚胎中具有空间限制性表达模式的转录本。我们的方法利用机器人自动化快速分析序列选择的cdna在全安装原位杂交测定。我们从胚胎cDNA文库中随机收集了778个不同的基因，确定了它们的空间分布，并表明其中很大一部分表现出表达的模式。此外，基因本体论研究揭示了具有共同表达模式的基因产物组，为在发育中起作用的大部分被忽视的效应分子提供了新的见解。正如本文所述，在原位对全载胚胎进行自动杂交证明是直接的，并为我们提供了一种非常有力的方法来全面调查早期胚胎的基因表达。从生物学意义的角度来看，我们发现许多空间限制性转录本对应于先前未在母体效应和合子致死性近乎饱和的遗传筛选中发现的新转录本的位点，这一发现尤其值得注意。
Here we describe a high-throughput screen to isolate transcripts with spatially restricted patterns of expression in early embryos. Our approach utilizes robotic automation for rapid analysis of sequence-selected cDNAs in a whole-mount in situ hybridization assay. We determined the spatial distribution of a random collection of 778 different genes from an embryonic cDNA library and show that a significant fraction of these exhibit patterned profiles of expression. In addition, gene ontology Studies revealed groups of gene products exhibiting shared expression patterns, providing new insights into the largely overlooked effector molecules that function in development. As described in this paper, automated hybridization to whole-mount embryos in Situ proved to be straightforward and provided us with a very powerful method for the global survey of gene expression in early embryos. From the perspective of biological significance, Our finding that many spatially restricted transcripts correspond to loci encoding novel transcripts that have not been previously identified in nearly saturating genetic screens for maternal effect and zygotic lethals is particularly notable.