Yeast Protocols

In this second edition of a widely used classic laboratory manual, leading experts utilize the tremendous progress and technological advances that have occurred to create a completely new collection of not only the major basic techniques, but also advanced protocols for yeast research and for using yeast as a host to study genes from other organisms. The authors provide detailed methods for the isolation of subcellular components-including organelles and macromolecules, for the basic cellular and molecular analysis specific for yeast cells, and for the creation of conditional mutant phenotypes that lend themselves to powerful genome manipulation. Additional protocols offer advanced approaches to study genetic interactions, DNA and chromatin metabolism, gene expression, as well as the foreign genes and gene products in yeast cells.

Basic Investigations in Saccharomyces cerevisiae.- Isolation of Nucleic Acids.- Purification of Yeast Peroxisomes.- Isolation of Yeast Plasma Membranes.- Isolation of Yeast Mitochondria.- Extraction of Yeast Lipids.- Two-Dimensional Gel Electrophoresis of Total Yeast Proteins.- Pulsed-Field Gel Electrophoresis of Budding Yeast Chromosomes.- Analysis of Yeast Lipids.- Yeast Fluorescence Microscopy.- Intracellular Expression of Recombinant Antibody Fluorescent Protein Fusions for Localization of Target Antigens in Schizosaccharomyces pombe.- Yeast Transformation by the LiAc/SS Carrier DNA/PEG Method.- Mutagenesis.- Gene Disruption in the Budding Yeast Saccharomyces cerevisiae.- Inducible Degron and Its Application to Creating Conditional Mutants.- Synthetic Lethal Screen.- Synthetic Genetic Array Analysis in Saccharomyces cerevisiae.- Two-Dimensional Agarose Gel Analysis of DNA Replication Intermediates.- Chromatin Assembly in a Crude Fraction From Yeast Cells.- Chromatin Immunoprecipitation to Study Protein-DNA Interactions in Budding Yeast.- Isolation of Yeast Nuclei and Micrococcal Nuclease Mapping of Nucleosome Positioning.- Study of Transcriptional Regulation Using a Reporter Gene Assay.- Assessing Telomeric Phenotypes.- Controlled Expression of Recombinant Genes and Preparation of Cell-Free Extracts in Yeast.- Production of Heterologous Proteins in Yeast With the Aid of the Hsp150? Carrier.- Yeast Two-Hybrid System Screening.- Import of Precursor Proteins Into Isolated Yeast Mitochondria.