Dupuy Denis, Li Qian-Ru, Deplancke Bart, Boxem Mike, Hao Tong, Lamesch Philippe, Sequerra Reynaldo, Bosak Stephanie, Doucette-Stamm Lynn, Hope Ian A, Hill David E, Walhout Albertha J M, Vidal Marc
Center for Cancer Systems Biology and Department of Cancer Biology, Dana-Farber Cancer Institute, Harvard Medical School, Boston, Massachusetts 02115, USA.
Genome Res. 2004 Oct;14(10B):2169-75. doi: 10.1101/gr.2497604.
An important aspect of the development of systems biology approaches in metazoans is the characterization of expression patterns of nearly all genes predicted from genome sequences. Such "localizome" maps should provide information on where (in what cells or tissues) and when (at what stage of development or under what conditions) genes are expressed. They should also indicate in what cellular compartments the corresponding proteins are localized. Caenorhabditis elegans is particularly suited for the development of a localizome map since all its 959 adult somatic cells can be visualized by microscopy, and its cell lineage has been completely described. Here we address one of the challenges of C. elegans localizome mapping projects: that of obtaining a genome-wide resource of C. elegans promoters needed to generate transgenic animals expressing localization markers such as the green fluorescent protein (GFP). To ensure high flexibility for future uses, we utilized the newly developed MultiSite Gateway system. We generated and validated "version 1.1" of the Promoterome: a resource of approximately 6000 C. elegans promoters. These promoters can be transferred easily into various Gateway Destination vectors to drive expression of markers such as GFP, alone (promoter::GFP constructs), or in fusion with protein-encoding open reading frames available in ORFeome resources (promoter::ORF::GFP).
后生动物系统生物学方法发展的一个重要方面是对从基因组序列预测的几乎所有基因的表达模式进行表征。这样的“定位组”图谱应提供有关基因在何处(何种细胞或组织中)以及何时(发育的哪个阶段或在何种条件下)表达的信息。它们还应指出相应蛋白质在哪些细胞区室中定位。秀丽隐杆线虫特别适合用于构建定位组图谱,因为其所有959个成体体细胞都可以通过显微镜观察到,并且其细胞谱系已被完整描述。在此,我们解决了秀丽隐杆线虫定位组图谱绘制项目面临的挑战之一:即获得用于生成表达诸如绿色荧光蛋白(GFP)等定位标记的转基因动物所需的全基因组秀丽隐杆线虫启动子资源。为确保未来使用具有高度灵活性,我们利用了新开发的多位点Gateway系统。我们生成并验证了“1.1版”启动子组:一个包含约6000个秀丽隐杆线虫启动子的资源。这些启动子可以轻松转移到各种Gateway目的载体中,以驱动诸如GFP等标记的表达,单独表达(启动子::GFP构建体),或与ORFeome资源中可用的蛋白质编码开放阅读框融合表达(启动子::ORF::GFP)。