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Functional profiling of the Saccharomyces cerevisiae genome

机译:酿酒酵母基因组的功能分析

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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.
机译:确定基因缺失的影响是了解基因功能的基本方法。常规的遗传筛选表现出偏见,并且常常会遗漏有助于表型的基因。我们系统地构建了酵母酿酒酵母的基因缺失突变体(注解的开放阅读框或ORF的96%)的近乎完整的集合。被称为“分子条形码”的DNA序列可以唯一地识别每个菌株,从而可以并行分析其生长,并可以通过与高密度寡核苷酸阵列杂交来定量评估每个基因的适应性贡献。我们显示了先前已知的新基因对于在六个充分研究的条件下最佳生长是必要的:高盐,山梨糖醇,半乳糖,pH 8,基本培养基和制霉菌素处理。在四种测试条件下,最佳生长还需要不到7%的信使RNA表达显着增加的基因。我们的结果验证了酵母基因缺失的收集作为功能基因组学的宝贵资源。

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