Patterned Gene Expression in Drosophila Development

描述（由申请人提供）：我们的长期目标是了解基因相互作用的复杂网络，它是正常发育、疾病和进化过程的基础。为此，我们需要确定一个生物体中全部基因的基因表达谱。在伯克利果蝇基因组计划（BDGP）中，我们建立了果蝇发育的基因表达资源，该资源包含空间和时间胚胎表达模式，以及基于基因本体（GO）使用标准化，受控词汇的模式注释。我们的数据库目前包含75,000张带注释的图像，这些图像显示了果蝇基因组中大约45%（6,000）的蛋白质编码基因的原位杂交产生的表达模式。空间表达数据与我们的发育微阵列数据集成。我们建议将这些研究扩展到：(1)剩余的55%的蛋白质编码基因，(2)选择性剪接转录物，(3)非编码RNA候选基因。生成RNA探针的主要资源是我们的果蝇基因收集（DGC），目前包含与80%的注释基因对应的cDNA克隆。为了捕获不具有代表性cDNA克隆（20%）的基因的表达模式，我们使用基因特异性PCR产物生成96孔格式的RNA探针。我们的研究产生的基因表达数据将为阐明果蝇中13664个基因的功能提供基础信息，并将有助于阐明包括人类在内的其他真核生物中同源基因的功能。此外，基因表达数据与基因、转录物和蛋白质序列的整合将促进发现调控相互作用网络的研究。使用标准化词汇可以跨物种描述基因表达和组织分化。阐明果蝇发育中全基因组基因表达和调控的机制将有助于理解人类组织的正常生长和分化，这是理解人类疾病的先决条件。

DESCRIPTION (provided by applicant): Our long-term objective is to understand the complex network of genetic interactions that underlies the processes of normal development, disease and evolution. For this, we need to determine gene expression profiles for the full complement of genes in an organism. At the Berkeley Drosophila Genome Project (BDGP), we have established a gene expression resource for Drosophila development that contains spatial and temporal embryonic expression patterns as well as annotations of the patterns using a standardized, controlled vocabulary, based on Gene Ontology (GO). Our database currently contains 75,000 annotated images showing expression patterns generated using in-situ hybridization of staged whole-mounted embryos for approximately 45% (6,000) of the protein-coding genes in the Drosophila genome. The spatial expression data are integrated with our developmental microarray data. We propose to extend these studies to: (1) the remaining 55% of the protein-coding genes, (2) alternatively-spliced transcripts, and (3) non-coding RNA candidate genes The primary resource for generation of RNA probes is our Drosophila Gene Collection (DGC) which currently contains cDNA clones corresponding to 80% of the annotated genes. To capture expression patterns for genes that do not have a representative cDNA clone (20%), we used gene-specific PCR products to generate RNA probes in 96-well format. The gene expression data produced by our study will provide fundamental information for elaborating the function of the 13,664 genes in Drosophila and will aid in elucidating the function of the homologous genes in other eukaryotes, including humans. In addition, the integration of the gene expression data with gene, transcript and protein sequences will promote research to discover networks of regulatory interactions. Using the standardized vocabulary allows cross-species descriptions of gene expression and tissue differentiation. Elucidating the mechanisms responsible for genome-wide gene expression and regulation in Drosophila development will aid in understanding normal growth and differentiation of tissues in humans, prerequisites for understanding human disease.