Abstract

The availability of complete genome sequences has created the need for comprehensive methods to explore gene function. S. cerevisiae is the first eukaryotic organism for which the DNA sequence became available, and exhibits many characteristics, such as amenable genetics and controllable physiology, which make it an ideal experimental system for the functional analysis of novel genes. The most direct way to determine gene function involves the disruption of the unknown genes followed by phenotypic analysis. In the last eight years, different systematic approaches have been employed to produce collections of mutant strains in which every putative ORF was erased from the yeast genome. Moreover, because of the high level of redundancy found in the yeast genome, new vectors and efficient methods have been created to perform multiple deletions in the same strain background. Such a collection of tools and mutants provide a major resource for both conventional functional analysis, and for the new whole-genome approaches aiming to characterise gene function via large-scale competition experiments.

Keywords: pcr-mediated gene replacement, functional analysis, systematic screen, genome, cre/loxp