Breslow David K, Cameron Dale M, Collins Sean R, Schuldiner Maya, Stewart-Ornstein Jacob, Newman Heather W, Braun Sigurd, Madhani Hiten D, Krogan Nevan J, Weissman Jonathan S
Department of Cellular and Molecular Pharmacology, University of California, San Francisco, 1700 4th Street, San Francisco, California 94158, USA.
Nat Methods. 2008 Aug;5(8):711-8. doi: 10.1038/nmeth.1234. Epub 2008 Jul 11.
Functional genomic studies in Saccharomyces cerevisiae have contributed enormously to our understanding of cellular processes. Their full potential, however, has been hampered by the limited availability of reagents to systematically study essential genes and the inability to quantify the small effects of most gene deletions on growth. Here we describe the construction of a library of hypomorphic alleles of essential genes and a high-throughput growth competition assay to measure fitness with unprecedented sensitivity. These tools dramatically increase the breadth and precision with which quantitative genetic analysis can be performed in yeast. We illustrate the value of these approaches by using genetic interactions to reveal new relationships between chromatin-modifying factors and to create a functional map of the proteasome. Finally, by measuring the fitness of strains in the yeast deletion library, we addressed an enigma regarding the apparent prevalence of gene dispensability and found that most genes do contribute to growth.
酿酒酵母中的功能基因组学研究极大地促进了我们对细胞过程的理解。然而，其全部潜力受到用于系统研究必需基因的试剂有限，以及无法量化大多数基因缺失对生长的微小影响的阻碍。在这里，我们描述了一个必需基因次等位基因突变体文库的构建，以及一种高通量生长竞争测定法，以以前所未有的灵敏度测量适应性。这些工具显著提高了在酵母中进行定量遗传分析的广度和精度。我们通过利用遗传相互作用揭示染色质修饰因子之间的新关系，并创建蛋白酶体的功能图谱，来说明这些方法的价值。最后，通过测量酵母缺失文库中菌株的适应性，我们解决了一个关于基因可 dispensability 明显流行的谜团，发现大多数基因确实对生长有贡献。
