High-throughput genetic interaction mapping in the fission yeast Schizosaccharomyces pombe
High-throughput genetic interaction mapping in the fission yeast Schizosaccharomyces pombe
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DOI：
10.1038/nmeth1098
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发表时间：
2007-10-01
期刊：
影响因子：
48
通讯作者：
Krogan, Nevan J.
中科院分区：
文献类型：
--
作者：
Roguev, Assen;Wiren, Marianna;Krogan, Nevan J.
上位分析报告了一个基因的功能在多大程度上依赖于另一个基因的存在，是研究细胞功能组织的有力工具。然而，上位性的系统全基因组研究受到限制，大部分数据收集于出芽酵母，酿酒酵母。在这里，我们提出了两种“pombe上位映射”策略，即PEM-1和PEM-2，它们允许在裂变酵母S. pombe中产生高通量双突变体。这些方法利用了先前描述的隐性环己亚胺抗性突变。这两种系统都可用于全基因组筛选或生成高密度、定量上位微型阵列谱（E-MAPs）。由于酿酒葡萄球菌和酿酒葡萄球菌的进化距离较远，该方法将提供对酿酒葡萄球菌中存在的保守生物学途径的深入了解，并将使遗传相互作用网络的保护得到全面分析。
Epistasis analysis, which reports on the extent to which the function of one gene depends on the presence of a second, is a powerful tool for studying the functional organization of the cell. Systematic genome-wide studies of epistasis, however, have been limited, with the majority of data being collected in the budding yeast, Saccharomyces cerevisiae. Here we present two 'pombe epistasis mapper' strategies, PEM-1 and PEM-2, which allow for high-throughput double mutant generation in the fission yeast, S. pombe. These approaches take advantage of a previously undescribed, recessive, cycloheximide-resistance mutation. Both systems can be used for genome-wide screens or for the generation of high-density, quantitative epistatic miniarray profiles ( E-MAPs). Since S. cerevisiae and S. pombe are evolutionary distant, this methodology will provide insight into conserved biological pathways that are present in S. pombe, but not S. cerevisiae, and will enable a comprehensive analysis of the conservation of genetic interaction networks.