A comprehensive strategy enabling high-resolution functional analysis of the yeast genome
A comprehensive strategy enabling high-resolution functional analysis of the yeast genome
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DOI：
10.1038/nmeth.1234
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发表时间：
2008-08-01
期刊：
影响因子：
48
通讯作者：
Weissman, Jonathan S.
中科院分区：
文献类型：
--
作者：
Breslow, David K.;Cameron, Dale M.;Weissman, Jonathan S.
酿酒酵母的功能基因组研究对我们对细胞过程的理解做出了巨大贡献。然而，由于系统研究必需基因的试剂有限，并且无法量化大多数基因缺失对生长的微小影响，它们的全部潜力受到了阻碍。在这里，我们描述了必需基因的亚等位基因库的构建和高通量生长竞争测定，以前所未有的灵敏度测量适应性。这些工具极大地提高了酵母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，我们解决了基因可分配性明显普遍存在的谜团，并发现大多数基因确实有助于生长。
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.