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- Title
A molecular barcoded yeast ORF library enables mode-of-action analysis of bioactive compounds.
- Authors
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
- Abstract
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.
- Subjects
YEAST fungi genetics; GENOMICS; BIOACTIVE compounds; DRUG resistance; PROMOTERS (Genetics); OLIGONUCLEOTIDES; BIOCHEMICAL genetics; DNA microarrays
- Publication
Nature Biotechnology, 2009, Vol 27, Issue 4, p369
- ISSN
1087-0156
- Publication type
Article
- DOI
10.1038/nbt.1534