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Includes bibliographical references and index.

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Tetrahymena thermophila is emerging as a powerful experimental system for functional genetics. The ciliated protozoan offers numerous advantages, not the least of which is the ability to eliminate any specific gene of interest and then to evaluate the effect of that mutation on the living cell. Past investigations with T. thermophila have yielded several key discoveries, including dynein, catalytic RNA, and telomerase. This volume is a comprehensive resource for using Tetrahymena in the laboratory and is aimed at persons already experienced, as well as newcomers to the organism. It covers both the biological background and essential protocols for investigators rapidly turning to Tetrahymena as the experimental system of choice. Key Features * Contains both theoretical and practical issues in 30 chapters contributed by the world authorities on Tetrahymena * Indispensible for both the novice and the experienced researcher * Overviews the history, cell biology, and genetics of the organism * Describes essential protocols on the growth of cells, genetic techniques, and how to look at the cells with the microscope * Illustrates how the methods can be applied to solve various cell biological problems * Reviews recently developed strategies for altering gene expression.

Front Cover; Methods in Cell Biology: Tetrahymena Thermophila; Copyright Page; Contents; Contributors; Preface; Part I: Overview; Chapter 1. Laboratory and Evolutionary History of Tetrahymena themophila; I. Characters in Search of a Script; II. Paramecium Precursors; III. Tetrahymena Breeding Studies; IV. Perils of Passage; V. Doing Genetics by Evolution; References; Chapter 2. Cell Biology of Tetrahymena thermophila; I. Introduction; II. Membranes and Membrane-Bounded Compartments; III. The Cytoskeleton and Membrane Skeleton; IV. Dynamics of the Cell Cycle; V. Spacial Patterning.

VI. Starvation-Mediated DevelopmentVII. Concluding Remarks; References; Chapter 3. Tetrahymena Genetics: Two Nuclei Are Better Than One; I. Asexual and Sexual Reproduction; II. Nuclear Dualism; III. Special Features of Tetrahymena Genetics; IV. DNA-Mediated Transformation; V. Summary and Prospects; References; Part II: Methods; Chapter 4. Tetrahymena as a Laboratory Organism: Useful Strains, Cell Culture, and Cell Line Maintenance; I. Introduction; II. Useful Tetrahymena Strains; III. Culture Media; IV. Culturing Tetrahymena Cells; V. Cell Line Storage; References.

Chapter 5. Long-Term StorageI. Introduction; II. Supplies; III. Preparation of Cells before Freezing; IV. Freezing; V. Thawing and Recovery; VI. Conclusion; References; Chapter 6. Genetic Crosses: Setting Up Crosses, Testing Progeny, and Isolating Phenotypic Assortants; I. Introduction; II. Methods for Mating Cells; III. How to Isolate and/or Select Progeny & amp;om a Cross; IV. Mating Type Tests and Sexual Immaturity Tests; V. Isolating Assortants; References; Chapter 7. Methods for Genetic Analysis; I. Introduction; II. Reagents; III. Equipment; IV. Strains; V. Strategies; References.

Chapter 8. Isolation of Micronuclear and Macronuclear DNAI. Introduction; II. Cell Culture; III. Percoll Method for Isolation of Nuclei; IV. Isolation of High Molecular Weight DNA from Nuclei; V. Solutions; References; Chapter 9. Genetically Sorting a Collection of Tetrahymena Mutants; I. Introduction; II. Mapping the Mutations to Chromosome Arms; III. Genetic Complementation Tests; References; Chapter 10. Genetically Mapping New Mutants and Cloned Genes; I. Introduction; II. Genetically Mapping a Mutation; III. Genetically Mapping a Cloned Gene or Sequence.

III. Special Procedures and EquipmentReferences; Chapter 11. Selection of Motility Mutants; I. Introduction; II. Materials; IV. Strains; V. Inducing and Screening for Temperature-Sensitive Mutations Affecting Cilia Regeneration; VI. Mutagenesis by Targeted Gene Knockout; References; Chapter 12. Nuclear and Cytoskeletal Fluorescence Microscopy Techniques; I. Introduction; II. DAPI: A Fluorescent Nuclear Stain; III. Immunofluorescence Labeling for Conventional Fluorescence Microscopy; IV. Immunofluorescence Labeling for Confocal Fluorescence Microscopy; References.

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