Universal Clone Resources for Drosophila Proteomics

描述（由申请人提供）：由于从多种生物体中产生大量基因组序列，基因组学的下一个主要挑战是鉴定和组装每种生物体的完整蛋白质组。在伯克利果蝇基因组计划（BDGP）中，我们正在积极地生成一个全面的cDNA资源，果蝇基因集（DGC）将包含约13，700个注释基因中的每一个的至少一个cDNA。我们的cDNA收集大大加快了对所有果蝇基因的全面转录图谱的进展，并被用于确定胚胎中的基因表达模式。目前，5，810个cDNA克隆包含DGC Gold Collection，其是一组无义和错义突变的全长cDNA。我们的目标是使用这些cDNA克隆构建两个集合的通用供体克隆，一个有和一个没有天然终止密码子。这些集合将是高质量开放阅读框架（ORF）的主要来源，其将容易转移到许多不同的表达载体中。此外，我们计划使用供体克隆制备两组表达克隆，一组用于组织培养细胞，另一组用于转基因果蝇。我们的目标是获得一个更详细的了解由果蝇基因组编码的蛋白质的完整集合，并为研究界提供功能基因组学和蛋白质组学的资源。这些克隆资源的构建将提供信息和工具，进一步加深我们对高等真核生物的理解，并为更完整地分析果蝇和其他真核生物（包括人类）中的蛋白质功能奠定基础。

DESCRIPTION (provided by applicant): As a result of the production of a vast amount of genomic sequence from a wide variety of organisms, the next major challenge in genomics is to identify and assemble complete protein sets for each. At the Berkeley Drosophila Genome Project (BDGP) we are actively generating a comprehensive cDNA resource, the Drosophila Gene Collection (DGC) that will contain at least one cDNA for each of the ~13,700 annotated genes. Our cDNA collection has greatly accelerated progress towards a comprehensive transcript map of all Drosophila genes and is being used to determine gene expression patterns in the embryo. Presently, 5,810 cDNA clones comprise the DGC Gold Collection, which is a set of full-length cDNAs that are free of nonsense and missense mutations. We aim to use these cDNA clones to construct two collections of universal donor clones, one with and one without the native stop codon. These collections will be the primary source of high quality open reading frames (ORFs) that will be easy to transfer into a number of different expression vectors. Further, we plan to use the donor clones to make two sets of expression clones, one for use in tissue culture cells and another for use in transgenic flies. Our goals are to obtain a more detailed understanding of the complete set of proteins that are encoded by the Drosophila melanogaster genome and to provide resources for functional genomics and proteomics to the research community. The construction of these clone resources will provide information and tools that will further our understanding of higher eukaryotes and lay the groundwork for more complete analyses of protein function in Drosophila and other eukaryotes, including humans.