Journal
NATURE BIOTECHNOLOGY
Volume 27, Issue 4, Pages 369-377Publisher
NATURE PUBLISHING GROUP
DOI: 10.1038/nbt.1534
Keywords
-
Categories
Funding
- NSERC
- NHGRI [MOP-84305, MOP-81340]
- NIGMS Center for Quantitative Biology [GM071508, R01 (GM046406)]
- National Institutes of Health [GM62637]
- Energy and Industrial Technology Development Organization (NEDO)
- Ontario Genomics Institute [2004-OGI-3-01]
- Canadian Institutes of Health [MOP-57830]
Ask authors/readers for more resources
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.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available