Anmerkung: Cover -- Copyright Page -- Contents -- Contributors -- Preface -- Part I: Overview -- Chapter 1. Laboratory and Evolutionary History of Tetrahymena thermophila -- Summary -- 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 Development -- VII. 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 Storage -- I. 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 from 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 DNA -- I. 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 -- References -- Chapter 11. Selection of Motility Mutants -- I. Introduction -- II. Materials -- III. Special Procedures and Equipment -- 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 -- Chapter 13. Nuclear and Cortical Histology for Brightfield Microscopy -- I. Introduction -- II. Silver Staining -- III. Protargol Staining -- IV. Giemsa Staining -- References -- Chapter 14. Fixation of Tetrahymena Cells for Electron Microscopy -- I. Introduction -- II. Cell Culture -- III. Fixation -- IV. Critical Point Drying for SEM -- V. Embedding for TEM -- VI. Staining Sections (TEM) -- References -- Chapter 15. Immunoelectron Microscopy of Tetrahymena -- I. Introduction -- II. General Considerations -- III. Postembedding Labeling -- IV. Double Labeling with the Postembedding Technique -- V. Pre-embedding Labeling -- VI. Controls for Immunogold Labeling. , VII. Quantitative Analysis of Colloidal Gold Labeling -- VIII. Troubleshooting: Problems and Recommended Remedies -- References -- Part III: Cell Biology -- Chapter 16. Regulated Protein Secretion in Tetrahymena thermophila -- I. Introduction -- II. Isolation of the Released Contents of Dense-Core Secretory Vesicles (Mucocysts) -- III. Stimulation of Cells to Test Exocytic Capability or to Study Postexocytic Events -- IV. Isolation of Intact Secretory Vesicles -- References -- Chapter 17. Electrophysiology of Tetrahymena -- I. Introduction -- II. Materials and Methods -- III. Commentary and Summary -- References -- Chapter 18. Isolation and Characterization of in Vivo Modified Histones and an Activity Gel Assay for Identification of Histone Acetyltransferases -- I. Introduction -- II. Methods -- References -- Chapter 19. Tetrahymena Telomerase Activity, Purification, and Reconstitution -- I. Introduction -- II. Materials and Methods -- III. Commentary and Summary -- References -- Chapter 20. Studying the Telomerase RNA in Tetrahymena -- I. Introduction -- II. General Strategy and Overview -- III. Basic Methods -- References -- Chapter 21. Isolation and Characterization of 22S Outer Arm Dynein from Tetrahymena Cilia -- I. Introduction -- II. Methods -- III. Summary -- References -- Chapter 22. Preparation of Cytoskeletal Fractions from Tetrahymena thermophila -- I. Introduction -- II. Methods -- References -- Chapter 23. Immunoprecipitation Procedures -- I. Introduction -- II. Method -- References -- Chapter 24. Tetrahymena Calcium-Binding Proteins, TCBP-25 and TCBP-23 -- I. Introduction -- II. Methods -- References -- Part IV: Manipulating Genes -- Chapter 25. Microinjection of Tetrahymena thermophila -- I. Introduction -- II. The Technique of Microinjection -- III. Concluding Remarks -- References. , Chapter 26. Transient and Stable DNA Transformation of Tetrahymena thermophila by Electroporation -- I. Introduction -- II. Considerations -- III. Preparing Plasmid DNA for Electrotransformation -- IV. General Protocol for Conjugant Electrotransformation -- V. Transformation with Integrating Vectors: Gene Replacements and Gene Knockouts -- VI. Transformation with Autonomously Replicating Vectors -- References -- Chapter 27. Biolistic Transformation of Macro- and Micronuclei -- I. Introduction -- II. Cells -- III. Transformation -- References -- Chapter 28. Knockout Heterokaryons Enable Facile Mutagenic Analysis of Essential Genes in Tetrahymena -- I. Introduction -- II. Strategy -- III. Methods for Creating Knockout Heterokaryons -- References -- Chapter 29. Creation and Use of Antisense Ribosomes in Tetrahymena thermophila -- I. Introduction -- II. Using Antisense Ribosomes in Tetrahymena thermophila -- III. Considerations Particular to Antisense Ribosomes -- IV. Evaluation and Applications of the Antisense Ribosome System -- References -- Chapter 30. Protein Tagging in Tetrahymena -- I. Introduction -- II. Strategies for Gene Tagging in Tetrahymena -- III. Methods -- References -- Appendix I. Genetic Nomenclature Rules for Tetrahymena thermophila -- Appendix II. Codon Usage in Tetrahymena thermophila -- Index -- Volumes in Series.