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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.
This little tale was written between two and three years ago, in the hope that it might help to call the attention of wiser and better men than I am, to the questions which are now agitating the minds of the rising generation, and to the absolute necessity of solving them at once and earnestly, unless we would see the faith of our forefathers crumble away beneath the combined influence of new truths which are fancied to be incompatible with it, and new mistakes as to its real essence. That this can be done I believe and know: if I had not believed it, I would never have put pen to paper on the subject.
Leland H. Hartwell Director, Fred Hutchinson Cancer Research Center, Nobel Laureate for Medicine, 2001 Yeast has proved to be the most useful single-celled organism for studying the fundamental aspects of cell biology. Resources are now available for yeast that greatly simplify and empower new investigations, like the presence of strains with each gene deleted, each protein tagged and databases on protein-protein interactions, gene regulation, and subcellular protein location. A powerful combination of genetics, cell biology, and biochemistry employed by thousands of yeast researchers has unraveled the complexities of numerous cellular processes from mitosis to secretion and even uncovered new insights into prion diseases and the role of prions in normal biology. These insights have proven, time and again, to foretell the roles of proteins and pathways in human cells. The collection of articles in this volume explores the use of yeast in pathway analysis and drug discovery. Yeast has, of course, supplied mankind's most ubiquitous drug for thousands of years. In one aspect, the role of yeast in drug discovery is much like the role of yeast in other areas of biology. Yeast offers the power of genetics and a repetoire of resources available in no other organism. Using yeast in the study of drug targets and metabolism can help to make a science of what has been largely an empirical activity. A science of drug discovery would permit rigorous answers to important questions.
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