A robust toolkit for functional profiling of the yeast genome
A robust toolkit for functional profiling of the yeast genome
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DOI：
10.1016/j.molcel.2004.09.035
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发表时间：
2004-11-05
期刊：
影响因子：
16
通讯作者：
Boeke, JD
中科院分区：
文献类型：
--
作者：
Pan, XW;Yuan, DS;Boeke, JD
突变表型的研究是理解基因功能的基本方法。构建近乎完整的酵母敲除 (YKO) 集合和识别每个菌株的独特分子条形码 (或 TAG) 可以对酿酒酵母进行定量功能分析。通过使用这些标签和 SGA 报告基因 MFA1pr-HIS3（促进杂合二倍体 YKO 菌株转化为单倍体突变体），我们开发了一套高效的基于微阵列的技术，统称为 dSLAM（基于微阵列的二倍体合成致死分析），以探测全基因组基因化学和基因基因 互动。直接比较表明，这些技术在酵母基因组功能分析方面比现有方法更可靠。这些工具的广泛应用将阐明全面的酵母遗传网络。
Study of mutant phenotypes is a fundamental method for understanding gene function. The construction of a near-complete collection of yeast knockouts (YKO) and the unique molecular barcodes (or TAGs) that identify each strain has enabled quantitative functional profiling of Saccharomyces cerevisiae. By using these TAGs and the SGA reporter, MFA1pr-HIS3, which facilitates conversion of heterozygous diploid YKO strains into haploid mutants, we have developed a set of highly efficient microarray-based techniques, collectively referred as dSLAM (diploid-based synthetic lethality analysis on microarrays), to probe genome-wide gene-chemical and gene-gene interactions. Direct comparison revealed that these techniques are more robust than existing methods in functional profiling of the yeast genome. Widespread application of these tools will elucidate a comprehensive yeast genetic network.