Fission Yeast: A Laboratory Manual

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Click to Enlarge				Fission Yeast: A Laboratory Manual Subject Area(s): Yeast; Cell Biology; Microbiology Edited by Iain Hagan, Cancer Research UK Manchester Institute, University of Manchester; Antony M. Carr, University of Sussex; Agnes Grallert, Cancer Research UK Manchester Institute, University of Manchester; Paul Nurse, The Francis Crick Institute *Download a Free Excerpt from Fission Yeast:* Preface Intro: Introduction to Fission Yeast as a Model System Chapter 2: Growth and the Environment of Schizosaccharomyces pombe Protocol: Mating, Spore Dissection, and Selection of Diploid Cells in Schizosaccharomyces japonicus Index

				© 2016 • 490 pages, illustrated (32 color, 17 B&W), index Hardcover • ~~$150~~ 120.00 ISBN 978-1-621820-81-9 You save: 20% You will receive free shipping on this item at checkout. Free shipping offer applies to direct website purchases by individual U.S. and Canada customers only. This title also available in: Paperback

Description Contents Related Titles Description Fission yeast are unicellular, rod-shaped fungi that divide by medial fission. Studies using fission yeast were instrumental in identifying fundamental mechanisms that govern cell division, differentiation, and epigenetics, to name but a few. Their rapid growth rate, genetic malleability, and similarities to more complex eukaryotes continue to make them excellent subjects for many biochemical, molecular, and cell biological studies. This laboratory manual provides an authoritative collection of core experimental procedures that underpin modern fission yeast research. The contributors describe basic methods for culturing and genetically manipulating fission yeast, synchronization strategies for probing the cell cycle, technologies for assessing proteins, metabolites, and cell wall constituents, imaging methods to visualize subcellular structures and dynamics, and protocols for investigating chromatin and nucleic acid metabolism. Modifications to techniques commonly used in related species (e.g., budding yeast) are noted, as are useful resources for fission yeast researchers, including various databases and repositories. The well-studied fission yeast Schizosaccharomyces pombe is the focus throughout, but the emerging model S. japonicus—a larger, dimorphic species with several desirable characteristics—is also covered. This manual is an important reference for existing fission yeast laboratories and will serve as an essential start-up guide for those working with fission yeast for the first time. Contents Preface CHAPTER 1 INTRODUCTION Introduction to Fission Yeast as a Model System Jacqueline Hayles and Paul Nurse CHAPTER 2 INTRODUCTION Growth and the Environment of Schizosaccharomyces pombe Janni Petersen and Paul Russell CHAPTER 3 INTRODUCTION Genetic Analysis of Schizosaccharomyces pombe Karl Ekwall and Geneviève Thon PROTOCOLS 1 Setting Up Schizosaccharomyces pombe Crosses/Matings Karl Ekwall and Geneviève Thon 2 Selecting Schizosaccharomyces pombe Diploids Karl Ekwall and Geneviève Thon 3 Spore Analysis and Tetrad Dissection of Schizosaccharomyces pombe Karl Ekwall and Geneviève Thon 4 Mating-Type Determination in Schizosaccharomyces pombe Karl Ekwall and Geneviève Thon 5 Ethyl Methanesulfonate Mutagenesis in Schizosaccharomyces pombe Karl Ekwall and Geneviève Thon CHAPTER 4 INTRODUCTION Molecular Genetic Tools and Techniques in Fission Yeast Johanne M. Murray, Adam T. Watson, and Antony M. Carr PROTOCOLS 1 Transformation of Schizosaccharomyces pombe: Electroporation Procedure Johanne M. Murray, Adam T. Watson, and Antony M. Carr 2 Transformation of Schizosaccharomyces pombe: Protoplast Procedure Johanne M. Murray, Adam T. Watson, and Antony M. Carr 3 Identifying Products of Recombinase-Mediated Cassette Exchange (RMCE) in Schizosaccharomyces pombe Johanne M. Murray, Adam T. Watson, and Antony M. Carr 4 Transformation of Schizosaccharomyces pombe: Lithium Acetate/ Dimethyl Sulfoxide Procedure Johanne M. Murray, Adam T. Watson, and Antony M. Carr 5 Extraction of Chromosomal DNA from Schizosaccharomyces pombe Johanne M. Murray, Adam T. Watson, and Antony M. Carr 6 Colony Polymerase Chain Reaction with Schizosaccharomyces pombe Johanne M. Murray, Adam T. Watson, and Antony M. Carr CHAPTER 5 INTRODUCTION Analysis of RNA Metabolism in Fission Yeast Jo Ann Wise and Olaf Nielsen PROTOCOLS 1 Preparation of Total RNA from Fission Yeast Jürg Bähler and Jo Ann Wise 2 Schizosaccharomyces pombe Polysome Profile Analysis and RNA Purification Dieter A. Wolf, Jürg Bähler, and Jo Ann Wise 3 4-Thiouridine Labeling to Analyze mRNA Turnover in Schizosaccharomyces pombe Juan Mata and Jo Ann Wise CHAPTER 6 INTRODUCTION Elementary Protein Analysis in Schizosaccharomyces pombe Agnes Grallert and Iain M. Hagan PROTOCOLS 1 Preparation of Protein Extracts from Schizosaccharomyces pombe Using Trichloroacetic Acid Precipitation Agnes Grallert and Iain M. Hagan 2 Small-Scale Immunoprecipitation from Fission Yeast Cell Extracts Agnes Grallert and Iain M. Hagan 3 Large-Scale Immunoprecipitation from Fission Yeast Cell Extracts Agnes Grallert and Iain M. Hagan 4 Large-Scale Purification of Small Ubiquitin-Like Modifier (SUMO)-Modified Proteins from Schizosaccharomyces pombe Minghua Nie and Michael N. Boddy 5 Improved Tandem Affinity Purification Tag and Methods for Isolation of Proteins and Protein Complexes from Schizosaccharomyces pombe Nicola Zilio and Michael N. Boddy CHAPTER 7 INTRODUCTION Stable Isotope Labeling by Amino Acids in Cell Culture (SILAC) Technology in Fission Yeast Boris Maček, Alejandro Carpy, André Koch, Claudia C. Bicho,  Weronika E. Borek, Silke Hauf, and Kenneth E. Sawin PROTOCOLS 1 Construction, Growth, and Harvesting of Fission Yeast Stable Isotope Labeling by Amino Acids in Cell Culture (SILAC) Strains André Koch, Claudia C. Bicho, Weronika E. Borek, Alejandro Carpy, Boris Maček, Silke Hauf, and Kenneth E. Sawin 2 Stable Isotope Labeling by Amino Acids in Cell Culture (SILAC)-Based Quantitative Proteomics and Phosphoproteomics in Fission Yeast Alejandro Carpy, André Koch, Claudia C. Bicho, Weronika E. Borek, Silke Hauf, Kenneth E. Sawin, and Boris Maček CHAPTER 8 INTRODUCTION Electron Microscopy of Fission Yeast J. Richard McIntosh, Mary K. Morphew, and Thomas H. Giddings, Jr. PROTOCOLS 1 Preparing Fission Yeast for Electron Microscopy Thomas H. Giddings, Jr., Mary K. Morphew, and J. Richard McIntosh 2 Immunolocalization of Proteins in Fission Yeast by Electron Microscopy Mary K. Morphew, Thomas H. Giddings, Jr., and J. Richard McIntosh 3 Cryoelectron Microscopy of Fission Yeast Mary K. Morphew, Thomas H. Giddings, Jr., and J. Richard McIntosh CHAPTER 9 INTRODUCTION Fixed-Cell Imaging of Schizosaccharomyces pombe Iain M. Hagan and Steven Bagley PROTOCOLS 1 Immunofluorescence Microscopy of Schizosaccharomyces pombe Using Chemical Fixation lain M. Hagan 2 Chromatin and Cell Wall Staining of Schizosaccharomyces pombe lain M. Hagan 3 Staining Fission Yeast Filamentous Actin with Fluorescent Phalloidin Conjugates lain M. Hagan CHAPTER 10 INTRODUCTION Live Cell Imaging in Fission Yeast Daniel P. Mulvihill PROTOCOLS 1 Microscopic Observation of Living Cells Stained with Fluorescent Probes Haruhiko Asakawa, Da-Qiao Ding, Tokuko Haraguchi, and  Yasushi Hiraoka 2 Visualization of a Specific Genome Locus by the lacO/Lacl-GFP System Da-Qiao Ding and Yasushi Hiraoka 3 Live Cell Imaging of the Schizosaccharomyces pombe Sexual Life Cycle Laura Merlini, Aleksandar Vjestica, Omaya Dudin, Felipe Bendezú, and Sophie G. Martin CHAPTER 11 INTRODUCTION Analysis of Schizosaccharomyces pombe Meiosis Akira Yamashita, Takeshi Sakuno, Yoshinori Watanabe, and  Masayuki Yamamoto PROTOCOLS 1 Live Imaging of Chromosome Segregation during Meiosis in the Fission Yeast Schizosaccharomyces pombe Akira Yamashita, Takeshi Sakuno, Yoshinori Watanabe, and Masayuki Yamamoto 2 Synchronous Induction of Meiosis in the Fission Yeast Schizosaccharomyces pombe Akira Yamashita, Takeshi Sakuno, Yoshinori Watanabe, and  Masayuki Yamamoto 3 A Simple Method to Induce Meiosis and Sporulation Semisynchronously in the Fission Yeast Schizosaccharomyces pombe Akira Yamashita, Takeshi Sakuno, Yoshinori Watanabe, and  Masayuki Yamamoto CHAPTER 12 INTRODUCTION Analysis of the Schizosaccharomyces pombe Cell Cycle lain M. Hagan, Agnes Grallert, and Viesturs Simanis PROTOCOLS 1 Cell Cycle Synchronization of Schizosaccharomyces pombe by Centrifugal Elutriation of Small Cells lain M. Hagan, Agnes Grallert, and Viesturs Simanis 2 Cell Cycle Synchronization of Schizosaccharomyces pombe by Lactose Gradient Centrifugation to Isolate Small Cells lain M. Hagan, Agnes Grallert, and Viesturs Simanis 3 Synchronizing Progression of Schizosaccharomyces pombe Cells from Prophase through Mitosis and into S Phase with nda3-KM311 Arrest Release lain M. Hagan, Agnes Grallert, and Viesturs Simanis 4 Synchronizing Progression of Schizosaccharomyces pombe Cells from G2 through Repeated Rounds of Mitosis and S Phase with cdc25-22 Arrest Release lain M. Hagan, Agnes Grallert, and Viesturs Simanis 5 Synchronization of S Phase in Schizosaccharomyces pombe Cells by Transient Exposure to M-Factor Pheromone Olaf Nielsen 6 Analyzing Schizosaccharomyces pombe DNA Content by Flow Cytometry Eric Boye, Silje Anda, Christiane Rothe, Trond Stokke, and Beata Grallert CHAPTER 13 INTRODUCTION Analysis of DNA Metabolism in Fission Yeast Francisco Antequera and Timothy C. Humphrey PROTOCOLS 1 Determination of the Frequency of Minichromosome Loss to Assess Chromosomal Stability in Fission Yeast Osami Niwa 2 Two-Dimensional Gel Electrophoresis of DNA Replication Intermediates in Schizosaccharomyces pombe Mar Sánchez and Francisco Antequera 3 Analysis of DNA Replication in Fission Yeast by Combing Divya R. lyer, Shankar Das, and Nick Rhind 4 Using Pulsed-Field Gel Electrophoresis to Analyze Schizosaccharomyces pombe Chromosomes and Chromosomal Elements Chen-Chun Pai, Carol Walker, and Timothy C. Humphrey 5 DNA Double-Strand Break Repair Assay Chen-Chun Pai, Elizabeth Blaikley, and Timothy C. Humphrey CHAPTER 14 INTRODUCTION Analysis of Heterochromatin in Schizosaccharomyces pombe Hugh P. Cam and Simon Whitehall PROTOCOLS 1 Reporter Gene Silencing Assays in Fission Yeast Hugh P. Cam and Simon Whitehall 2 Analysis of Schizosaccharomyces pombe Heterochromatin-Associated Short Interfering RNAs Ke Zhang 3 Micrococcal Nuclease Digestion of Schizosaccharomyces pombe Chromatin Hugh P. Cam and Simon Whitehall 4 Chromatin Immunoprecipitation (ChIP) in Schizosaccharomyces pombe Hugh P. Cam and Simon Whitehall CHAPTER 15 INTRODUCTION Fission Yeast Cell Wall Analysis Pilar Pérez and Juan C. Ribas PROTOCOL 1 Radioactive Labeling and Fractionation of Fission Yeast Walls Pilar Pérez and Juan C. Ribas CHAPTER 16 INTRODUCTION High-Throughput Quantitative Genetic Interaction Mapping in the Fission Yeast Schizosaccharomyces pombe Assen Roguev, Colm J. Ryan, Edgar Hartsuiker, and Nevan J. Krogan PROTOCOLS 1 Transformation of Schizosaccharomyces pombe in a 96-Well Format Assen Roguev, Jiewei Xu, and Nevan J. Krogan 2 DNA Preparation from Schizosaccharomyces pombe Assen Roguev, Jiewei Xu, and Nevan J. Krogan 3 Genetic Interaction Screens in Schizosaccharomyces pombe Using the Pombe Epistasis Mapper (PEM) System and a Manual Colony Replicator Isabelle Colson and Edgar Hartsuiker 4 Genetic Interaction Mapping in Schizosaccharomyces pombe Using the Pombe Epistasis Mapper (PEM) System and a ROTOR HDA Colony Replicating Robot in a 1536 Array Format Assen Roguev, Jiewei Xu, and Nevan J. Krogan 5 Genetic Interaction Score (S-Score) Calculation, Clustering, and Visualization of Genetic Interaction Profiles for Yeast Assen Roguev, Colm J. Ryan, Jiewei Xu, Isabelle Colson, Edgar Hartsuiker, and Nevan J. Krogan CHAPTER 17 INTRODUCTION Metabolomic Analysis of Schizosaccharomyces pombe: Sample Preparation, Detection, and Data Interpretation Tomáš Pluskal and Mitsuhiro Yanagida PROTOCOLS 1 Preparation of Intracellular Metabolite Extracts from Liquid Schizosaccharomyces pombe Cultures Tomáš Pluskal, Takahiro Nakamura, and Mitsuhiro Yanagida 2 Measurement of Metabolome Samples Using Liquid Chromatography-Mass Spectrometry, Data Acquisition, and Processing Tomas Pluskal and Mitsuhiro Yanagida CHAPTER 18 INTRODUCTION Schizosaccharomyces japonicus: A Distinct Dimorphic Yeast among the Fission Yeast Keita Aoki, Kanji Furuya, and Hironori Niki PROTOCOLS 1 Mating, Spore Dissection, and Selection of Diploid Cells in Schizosaccharomyces japonicus Kanji Furuya and Hironori Niki 2 Transformation of Schizosaccharomyces japonicus Keita Aoki and Hironori Niki 3 Induction of Hyphal Growth in Schizosaccharomyces japonicus Hironori Niki APPENDIX General Safety and Hazardous Material Information INDEX Related Titles Fission Yeast: A Laboratory Manual [Paperback]