Yeast Protocols

About this book

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.

Contents

Preface; Contributors; 1. Basic Investigations in Saccharomyces cerevisiae; Brendan Curran and Virginia Bugeja; 2. Isolation of Nucleic Acids; Michelle Hanna and Wei Xiao; 3. Purification of Yeast Peroxisomes; Ben Distal and Astrid Kragt; 4. Isolation of Yeast Plasma Membranes; Barry Panaretou and Peter Piper; 5. Isolation of Yeast Mitochondria; Chris Meisinger, Nikolaus Pfanner and Kaye N. Truscott; 6. Extraction of Yeast Lipids; Roger Schneiter and Gunther Daum; 7. Two-Dimensional Gel Electrophoresis of Total Yeast Proteins; Helian Boucherie and Christelle Monribot-Espagne; 8. Pulsed Field Gel Electrophoresis of Budding Yeast Chromosomes; Laura Maringele and David Lydall; 9. Analysis of Yeast Lipids; Roger Schneiter and Gunther Daum; 10. Yeast Fluorescence Microscopy; Jiri Hasek; 11. Intracellular Expression of Recombinant Antibody Fluorescent Protein Fusions for Localization of Target Antigens in Schizosaccharomyces pombe; Michelle A. Alting-Mees, Eddy P. Risseeuw, Enwu Liu, Michel Desautels, William L. Crosby, and Sean M. Hemmingsen; 12. Yeast Transformed by the LiAc/SS Carrier DNA/PEG Method; R. Daniel Gietz and Robin A. Woods; 13. Mutagenesis; Leslie Barbour, Michelle Hanna, and Wei Xiao; 14. Gene Disruption in the Budding Yeast Saccharomyces cerevisiae; Johannes H. Hegemann, Ulrich Guldener, and Gabriele J. Kohler. 15. Inducible Degron and Its Application to Creating Conditional Mutants; R. Jurgen Dohmen; 16. Synthetic Lethal Screen; Leslie Barbour and Wei Xiao; 17. Synthetic Genetic Array Analysis in Saccharomyces cerevisiae; Amy Hin Yan Tong and Charles Boone; 18. Two-Dimensional Agarose Gel Analysis of DNA Replication Intermediates; Alain T. Dandjinou, Michel Larrivee, Ralf E. Wellinger, and Raymund J. Wellinger; 19. Chromatin Assembly in a Crude Fraction From Yeast Cells; Karen M. Robinson and Michael C. Schultz; 20. Chromatin Immunoprecipitation to Study Protein-DNA Interactions in Budding Yeast; Elena Ezhkova and William P. Tansey; 21. Isolation of Yeast Nuclei and Micrococcal Nuclease Mapping of Nucleosome Positioning; Zhengjian Zhang and Joseph C. Reese; 22. Study of Transcriptional Regulation Using a Reporter Gene Assay; Yu Fu and Wei Xiao; 23. Assessing Telomeric Phenotypes; Catherine LeBel, Michel Larrivee, Amadou Bah, Nancy Laterreur, Nancy Levesque, and Raymund J. Wellinger; 24. Controlled Expression of Recombinant Genes and Preparation of Cell-Free Extracts in Yeast; Zhigang Wang; 25. Production of Heterologous Proteins in Yeast With The Aid of The Hsp 150[DELTA] Carrier; Marja Makarow, Anna-Liisa Hanninen, Taina Suntio, and Ricardo Nunes Bastos; 26. Yeast Two-Hybrid System Screening; R. Daniel Gietz.; 27. Import of Precursor Proteins into Isolated Yeast Mitochondria; Nils Wiedemann, Nikolaus Pfanner, and Peter Rehling.

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