title: Competition experiments coupled with high-throughput analyses for functional genomics studies in yeast.
abstract: Competition experiments are an effective way to provide a measurement of the fitness of yeast strains. The availability of the Saccharomyces cerevisiae yeast knock-out (YKO) deletion collection allows scientists to retrieve fitness data for the ~6,000 S. cerevisiae genes at the same time in a given environment. The molecular barcodes, characterizing each yeast mutant, serve as strain identifiers, which can be detected in a single microarray analysis. Competition experiments in continuous culture using chemically defined media allow a more specific discrimination of the strains based on their fitness profile. With this high-throughput approach, a series of genes that, when one allele is missing, result in either defective (haplo-insufficient) or favored (haplo-proficient) growth phenotype have been discovered, for each nutrient-limiting condition tested. While haplo-insufficient genes seemed to overlap largely across all the media used, the haplo-proficient ones seem to be more environment specific. For example, genes involved in the protein secretion pathway were highly haplo-insufficient in all the contexts, whereas most of the genes encoding for proteasome components showed a haplo-proficient phenotype specific to nitrogen-limiting conditions. In this chapter, the method used for implementation of competition experiments for high-throughput studies in yeast is presented.
authors.affiliation: Faculty of Life Sciences, The University of Manchester, Manchester, UK. D.Delneri@manchester.ac.uk
authors.forename: Daniela
authors.initials: D
authors.lastname: Delneri
mesh_terms: D061249:Batch Cell Culture Techniques; D003470:Culture Media; D046508:Culture Techniques / Q000295:instrumentation / Q000379:methods*; D005285:Fermentation; D023281:Genomics / Q000379:methods*; D009154:Mutation; D021141:Nucleic Acid Amplification Techniques; D009693:Nucleic Acid Hybridization; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000254:growth & development* / Q000302:isolation & purification / Q000378:metabolism
chemical_list: D003470:Culture Media