A molecular barcoded yeast ORF library enables mode-of-action analysis of bioactive compounds

作者：Ho Cheuk Hei; Magtanong Leslie; Barker Sarah L; Gresham David; Nishimura Shinichi; Natarajan Paramasivam; Koh Judice L Y; Porter Justin; Gray Christopher A; Andersen Raymond J; Giaever Guri; Nislow Corey; Andrews Brenda; Botstein David; Graham Todd R; Yoshida Minoru; Boone Charles^*

来源：Nature Biotechnology, 2009, 27(4): 369-377.

DOI：10.1038/nbt.1534

We present a yeast chemical-genomics approach designed to identify genes that when mutated confer drug resistance, thereby providing insight about the modes of action of compounds. We developed a molecular barcoded yeast open reading frame (MoBY-ORF) library in which each gene, controlled by its native promoter and terminator, is cloned into a centromere-based vector along with two unique oligonucleotide barcodes. The MoBY-ORF resource has numerous genetic and chemical-genetic applications, but here we focus on cloning wild-type versions of mutant drug-resistance genes using a complementation strategy and on simultaneously assaying the fitness of all transformants with barcode microarrays. The complementation cloning was validated by mutation detection using whole-genome yeast tiling microarrays, which identified unique polymorphisms associated with a drug-resistant mutant. We used the MoBY-ORF library to identify the genetic basis of several drug-resistant mutants and in this analysis discovered a new class of sterol-binding compounds.

• 出版日期2009-4