Functional profiling of the Saccharomyces cerevisiae genome
Functional profiling of the Saccharomyces cerevisiae genome
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DOI：
10.1038/nature00935
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发表时间：
2002-07-25
期刊：
影响因子：
64.8
通讯作者：
Johnston, M
中科院分区：
文献类型：
--
作者：
Giaever, G;Chu, AM;Johnston, M
确定基因缺失的影响是理解基因功能的一种基本方法。传统的遗传筛选存在偏差，对表型有贡献的基因常常被遗漏。我们系统性地构建了一个近乎完整的酿酒酵母（Saccharomyces cerevisiae）基因缺失突变体集合（96%的带注释的开放阅读框，即ORFs）。被称为“分子条形码”的DNA序列能够唯一地识别每个菌株，从而能够对它们的生长进行平行分析，并通过与高密度寡核苷酸阵列杂交来定量评估每个基因对适应性的贡献。我们表明，在六种经过充分研究的条件下，即高盐、山梨醇、半乳糖、pH 8、基本培养基和制霉菌素处理，先前已知的和新的基因对于最佳生长是必需的。在四种测试条件下，信使RNA表达显著增加的基因中，只有不到7%对于最佳生长也是必需的。我们的结果验证了酵母基因缺失集合是功能基因组学的一种宝贵资源。
Determining the effect of gene deletion is a fundamental approach to understanding gene function. Conventional genetic screens exhibit biases, and genes contributing to a phenotype are often missed. We systematically constructed a nearly complete collection of gene-deletion mutants (96% of annotated open reading frames, or ORFs) of the yeast Saccharomyces cerevisiae. DNA sequences dubbed 'molecular bar codes' uniquely identify each strain, enabling their growth to be analysed in parallel and the fitness contribution of each gene to be quantitatively assessed by hybridization to high-density oligonucleotide arrays. We show that previously known and new genes are necessary for optimal growth under six well-studied conditions: high salt, sorbitol, galactose, pH 8, minimal medium and nystatin treatment. Less than 7% of genes that exhibit a significant increase in messenger RNA expression are also required for optimal growth in four of the tested conditions. Our results validate the yeast gene-deletion collection as a valuable resource for functional genomics.