PCR-based engineering of yeast genome

Abstract

Publisher Summary The use of Saccharomyces cerevisiae as a model organism continues to increase as more genome sequences from higher eukaryotes become available and investigators who have previously concentrated on studying genes in larger eukaryotes now address the function of potential yeast homologs. High rates of homologous recombination in Saccharomyces cerevisiae allow precisely targeted modification of specific yeast chromosomal genes, an attractive feature of this model organism. Single-step PCR-based methods have been developed that allow more facile and economical gene deletion, epitope and/or protein tagging, or promoter substitution than traditional methods. Homologous recombination is commonly used to target modification of the yeast genome. Often one of the first steps in studying a yeast gene is to delete the gene using homologous recombination and to investigate the phenotype of the resuiting strain. Other targeted modifications include placing genes under the control of heterologous regulatable or constitutive promoters, introducing epitope/protein tags to the target protein, or introducing a mutation into the target gene.