Donnelly Centre for Cellular and Biomolecular Research, University of Toronto, 160 College Street, Toronto, ON M5S 3E1, Canada.
Nucleic Acids Res. 2012 Jan;40(Database issue):D687-94. doi: 10.1093/nar/gkr827. Epub 2011 Oct 18.
About one-fifth of the genes in the budding yeast are essential for haploid viability and cannot be functionally assessed using standard genetic approaches such as gene deletion. To facilitate genetic analysis of essential genes, we and others have assembled collections of yeast strains expressing temperature-sensitive (ts) alleles of essential genes. To explore the phenotypes caused by essential gene mutation we used a panel of genetically engineered fluorescent markers to explore the morphology of cells in the ts strain collection using high-throughput microscopy. Here, we describe the design and implementation of an online database, PhenoM (Phenomics of yeast Mutants), for storing, retrieving, visualizing and data mining the quantitative single-cell measurements extracted from micrographs of the ts mutant cells. PhenoM allows users to rapidly search and retrieve raw images and their quantified morphological data for genes of interest. The database also provides several data-mining tools, including a PhenoBlast module for phenotypic comparison between mutant strains and a Gene Ontology module for functional enrichment analysis of gene sets showing similar morphological alterations. The current PhenoM version 1.0 contains 78,194 morphological images and 1,909,914 cells covering six subcellular compartments or structures for 775 ts alleles spanning 491 essential genes. PhenoM is freely available at http://phenom.ccbr.utoronto.ca/.
大约五分之一的酿酒酵母基因对于单倍体的存活是必需的,并且不能使用标准的遗传方法(如基因缺失)进行功能评估。为了方便对必需基因进行遗传分析,我们和其他人一起构建了一系列表达温度敏感(ts)等位基因的酿酒酵母菌株的集合。为了探索必需基因突变引起的表型,我们使用一组经过基因工程改造的荧光标记物,通过高通量显微镜探索 ts 菌株集合中细胞的形态。在这里,我们描述了一个在线数据库 PhenoM(酵母突变体的表型组学)的设计和实现,用于存储、检索、可视化和挖掘从 ts 突变细胞的显微照片中提取的定量单细胞测量数据。PhenoM 允许用户快速搜索和检索感兴趣基因的原始图像及其定量形态数据。该数据库还提供了几个数据挖掘工具,包括用于比较突变株之间表型的 PhenoBlast 模块和用于功能富集分析显示相似形态改变的基因集的基因本体论模块。当前的 PhenoM 版本 1.0 包含 78194 个形态图像和 1909914 个细胞,涵盖六个亚细胞区室或结构,用于 775 个 ts 等位基因,跨越 491 个必需基因。PhenoM 可在 http://phenom.ccbr.utoronto.ca/ 免费获取。