eSGA:: E. coli synthetic genetic array analysis
eSGA:: E. coli synthetic genetic array analysis
复制标题
DOI：
10.1038/nmeth.1239
复制
发表时间：
2008-09-01
期刊：
影响因子：
48
通讯作者：
Emili, Andrew
中科院分区：
文献类型：
--
作者：
Butland, Gareth;Babu, Mohan;Emili, Andrew
物理和功能相互作用决定了细胞的分子组织。遗传相互作用，或上位性，往往发生在参与平行途径或相互关联的生物过程的基因产物之间。已经为酿酒酵母、粟酒裂殖酵母、秀丽隐杆线虫和黑腹果蝇设计了在全基因组范围内检测遗传相互作用的高通量实验系统，但此前尚未有原核生物的相关报道。在此，我们描述了一种定量筛选方法的开发，该方法用于监测细菌的遗传相互作用，其基于大肠杆菌缺失或亚效等位基因菌株的接合，以在全基因组范围内创建双突变体。我们观察到的某些双突变体组合的合成致病和合成致死（加重的遗传相互作用）模式提供了有关途径之间功能关系和冗余的信息，并使我们能够将细菌基因产物分组为功能模块。
Physical and functional interactions define the molecular organization of the cell. Genetic interactions, or epistasis, tend to occur between gene products involved in parallel pathways or interlinked biological processes. High-throughput experimental systems to examine genetic interactions on a genome-wide scale have been devised for Saccharomyces cerevisiae, Schizosaccharomyces pombe, Caenorhabditis elegans and Drosophila melanogaster, but have not been reported previously for prokaryotes. Here we describe the development of a quantitative screening procedure for monitoring bacterial genetic interactions based on conjugation of Escherichia coli deletion or hypomorphic strains to create double mutants on a genome-wide scale. The patterns of synthetic sickness and synthetic lethality (aggravating genetic interactions) we observed for certain double mutant combinations provided information about functional relationships and redundancy between pathways and enabled us to group bacterial gene products into functional modules.