Large-scale analysis of the yeast genome by transposon tagging and gene disruption
Large-scale analysis of the yeast genome by transposon tagging and gene disruption
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DOI：
10.1038/46558
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发表时间：
1999-11-25
期刊：
影响因子：
64.8
通讯作者：
Snyder, M
中科院分区：
文献类型：
--
作者：
Ross-Macdonald, P;Coelho, PSR;Snyder, M
在基因组规模上分析基因功能的经济方法相对较少。为了满足这一需求，我们已经开发了一种转座子标签策略的全基因组分析的破坏表型，基因表达和蛋白质定位，并已将这种方法应用于大规模的基因功能分析在芽殖酵母酿酒酵母。在这里，我们提出了有史以来最大的集合定义的酵母突变体在一个单一的遗传背景下产生的收集超过11,000株，每个携带转座子插入在营养生长和/或孢子形成过程中表达的基因组区域。这些插入影响了近2，000个注释基因，占酵母基因组中6，200个预测基因的约三分之一（1，2）。我们已经使用这个集合来确定使用20种不同生长条件的近8，000种菌株的破坏表型;将所得数据集进行聚类以识别功能相关基因的组。我们还确定了超过300个以前未注释的开放阅读框架，并通过间接免疫荧光分析了超过1,300个转座子标记的蛋白质。总的来说，我们的研究包括超过260，000个数据点，构成了有史以来最大的酵母基因组功能分析。
Economical methods by which gene function maybe analysed on a genomic scale are relatively scarce. To fill this need, we have developed a transposon-tagging strategy for the genome-wide analysis of disruption phenotypes, gene expression and protein localization, and have applied this method to the large-scale analysis of gene function in the budding yeast Saccharomyces cerevisiae. Here we present the largest collection of defined yeast mutants ever generated within a single genetic background-a collection of over 11,000 strains, each carrying a transposon inserted within a region of the genome expressed during vegetative growth and/or sporulation. These insertions affect nearly 2,000 annotated genes, representing about one-third of the 6,200 predicted genes in the yeast genome(1,2). We have used this collection to determine disruption phenotypes for nearly 8,000 strains using 20 different growth conditions; the resulting data sets were clustered to identify groups of functionally related genes. We have also identified over 300 previously non-annotated open reading frames and analysed by indirect immunofluorescence over 1,300 transposon-tagged proteins. In total, our study encompasses over 260,000 data points, constituting the largest functional analysis of the yeast genome ever undertaken.