Methods for large-scale analysis of gene expression, protein localization, and disruption phenotypes in Saccharomyces cerevisiae

Using a yeast genomic library mutagenized by transposon insertion, a bank of yeast strains containing random lacZ insertions throughout the genome can be generated. Such strains can be analyzed for gene expression, localization of beta-galactosidase-fusion proteins, and disruption phenotypes. The library can be hence be utilized for a variety of purposes, including insertional mutagenesis, analysis of differential gene expression, and identification of downstream targets of a protein of interest. We describe techniques for generating and utilizing such a bank and protocols we have developed to allow these analyses to be performed rapidly on a large number of individual strains. Also included are modifications of existing protocols for yeast transformation and DNA isolation to allow their use on large numbers of samples. (C) 1996 Wiley-Liss, Inc.