用于酵母基因组功能分析的强大工具集。

A robust toolkit for functional profiling of the yeast genome.

作者信息

Pan Xuewen, Yuan Daniel S, Xiang Dong, Wang Xiaoling, Sookhai-Mahadeo Sharon, Bader Joel S, Hieter Philip, Spencer Forrest, Boeke Jef D

机构信息

Department of Molecular Biology and Genetics, The Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.

出版信息

Mol Cell. 2004 Nov 5;16(3):487-96. doi: 10.1016/j.molcel.2004.09.035.

DOI:10.1016/j.molcel.2004.09.035

PMID:15525520

Abstract

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.

摘要

对突变体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基因敲除文库（YKO）以及用于识别每个菌株的独特分子条形码（或标签），使得对酿酒酵母进行定量功能分析成为可能。通过使用这些标签和SGA报告基因MFA1pr-HIS3（其有助于将杂合二倍体YKO菌株转化为单倍体突变体），我们开发了一套基于微阵列的高效技术，统称为dSLAM（基于微阵列的二倍体合成致死分析），以探究全基因组范围的基因-化学和基因-基因相互作用。直接比较表明，这些技术在酵母基因组功能分析中比现有方法更可靠。这些工具的广泛应用将阐明一个全面的酵母遗传网络。