An Updated Collection of Sequence Barcoded Temperature-Sensitive Alleles of Yeast Essential Genes.
An Updated Collection of Sequence Barcoded Temperature-Sensitive Alleles of Yeast Essential Genes.
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酵母基本基因的序列序列温度敏感等位基因的更新集合。
DOI：
10.1534/g3.115.019174
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发表时间：
2015-07-14
期刊：
影响因子：
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通讯作者：
Stirling PC
中科院分区：
文献类型：
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作者：
Kofoed M;Milbury KL;Chiang JH;Sinha S;Ben-Aroya S;Giaever G;Nislow C;Hieter P;Stirling PC
已经使用杂合二倍体、启动子关闭等位基因、通过具有不稳定的mRNA、不稳定的蛋白质或携带导致温度敏感（ts）表型的突变的等位基因的集合，在酿酒酵母中使用突变体集合进行了必需基因功能的系统分析。我们先前描述了以系统方式构建条形码化ts等位基因的方法。在这里，我们报告完成了这个收集的等位基因，涵盖600个必要的酵母基因。该资源涵盖了比以前的集合更大的基因库，并提供了一套适合于单基因和基因组分析的互补菌株。我们使用深度测序来表征在一半等位基因中导致ts表型的氨基酸变化。我们还使用高通量方法来描述等位基因的相对ts行为。最后，我们证明了实验有用的收集在一个高内容，功能基因组筛选TS等位基因，增加自发P-体形成。通过增加等位基因的数量和改进注释，这个ts集合将作为探索必需酵母基因生物学新方面的社区资源。
Systematic analyses of essential gene function using mutant collections in Saccharomyces cerevisiae have been conducted using collections of heterozygous diploids, promoter shut-off alleles, through alleles with destabilized mRNA, destabilized protein, or bearing mutations that lead to a temperature-sensitive (ts) phenotype. We previously described a method for construction of barcoded ts alleles in a systematic fashion. Here we report the completion of this collection of alleles covering 600 essential yeast genes. This resource covers a larger gene repertoire than previous collections and provides a complementary set of strains suitable for single gene and genomic analyses. We use deep sequencing to characterize the amino acid changes leading to the ts phenotype in half of the alleles. We also use high-throughput approaches to describe the relative ts behavior of the alleles. Finally, we demonstrate the experimental usefulness of the collection in a high-content, functional genomic screen for ts alleles that increase spontaneous P-body formation. By increasing the number of alleles and improving the annotation, this ts collection will serve as a community resource for probing new aspects of biology for essential yeast genes.