Methods in Cell Biology

Name: Methods in Cell Biology
Brand: Academic Press
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Academic Press

Published on 23. October 1978

519 pages

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978-0-08-085917-0 (ISBN)

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Front Cover
Methods in Cell Biology, Volume XX
Copyright Page
Contents
List of Contributors
Preface
Chapter 1. Measuring Spontaneous Mutation Rates in Yeast
I. Introduction
II. Measuring the Spontaneous Mutation Rate in Mitotic Cells by Limiting a Required Nutrient
III. Measuring the Spontaneous Mutation Rate in Mitotic Cells Grown in Complete Medium
IV. Measuring the Spontaneous Mutation Rate during Meiosis
V. Conclusion
Appendix: Biometrical Aspects of Measuring Mutation Rates
References
Chapter 2. Manganese Mutagenesis in Yeast
I. Introduction
II. Induction of Mitochondrial Mutations
III. Induction of Nuclear Mutations
IV. The Uptake of Manganese and Its General Effects on Yeast Cells
V. Other Mutagens Inducing Mutations Nonlethal for rho Factor
References
Chapter 3. Yeast Cell Selection and Synchrony: Density Gradients and Mating Factor Block
I. Introduction
II. lsopycnic Density Gradients
III. Isolation of Cells Labeled at Specific Periods of the Cell Cycle
IV. a- and a-Factor Selection and Synchronization of Haploid Yeast Cells
V. Discussion
References
Chapter 4. The Ribosomal Proteins of Saccharomyces cerevisiae
I. Preparation of Ribosomes
II. Preparation of Ribosomal Proteins
III. Analysis of Ribosomal Proteins
IV. Conclusion
References
Note Added in Proof
Chapter 5. Isolation, Characterization, and Translation of mRNA from Yeast
I. Media, Cells, and Spheroplasts
II. Isolation of Polysomes, Single Ribosomes, and Ribosomal Subunits
III. Detection of mRNA in Polysomes and Subunits
IV. Preparation of RNA
V. Analysis of RNA on Polyacrylamide Gels
VI. Isolation of Poly-A-Containing RNA
VII. Analysis of 5'-Termini of mRNA
VIII. Translation of Yeast mRNA
References
Chapter 6. Method for Yeast Protein Synthesis in a Cell-Free Systern
I. Introduction
II. Early Yeast Cell-Free Systems
III. The S-30 Extract
IV. Assay of Protein Synthesis
V. Prospects
References
Chapeter 7. Preparation of Protoplasts of Schizosaccharomyces pombe
I. Introduction
II. Preparation of Protoplasts
III. Concluding Remarks
References
Chapter 8. Bulk Isolation of Yeast Mitochondria
I. Introduction
II. Materials and Methods
III.Breakage of Yeast Cells
IV.Isolation of Mitochondria
V.Properties of Isolated Mitochondria
VI.Discussion
References
Chapter 9. Isolation, Regeneration, and Fusion of Protoplasts of Phycomyces
I. Introduction
II. Material
III. Preparation of Cells
IV.Protoplast Isolation
V.Reversion of Protoplasts
VI.Protoplast Fusion
VII.Discussion
References
Chapter 10.The Neurospora Plasma Membrane: A New Experimental System for Investigating Eukaryote Surface Membrane Structure and Function
I. Introduction
II.Isolation of Neurospora Plasma Membranes
III.Properties of Isolated Neurospora Plasma Membranes
IV.Conclusion
References
Chapter 11. Staining Fungal Nuclei with Mithramycin
I.Introduction
II.Mithramycin
III.Nuclear Staining Procedures
IV.Microscopy
V.Summary
References
Chapeter 12. Methods for Protoplast Formation in Escherichia coli
I. Introduction
II.General Methods
Ill.Appearance of Protoplasts
IV.Discussion
V.Summary and Conclusion
References
Chapter 13. Fusion of Bacterial Protoplasts
I. Background
II. Rationale for Fusion Experiment
III.Procedures for Bacterial Fusion
IV.Explored Variations and Alternatives
V.Criteria for Demonstration of Fusion
References
Chapter 14. Preparation of Protoplasts of Plant Cells
I.Introduction
II.Protoplast Isolation Procedures
III.Discussion
IV.Conclusions
References
Chapter 15. The Cultivation of Animal Cells and Production of Viruses in Serum-Free Systems
I. Introduction-The Need for Serum-Free Systems
II. The Role of Serum
III.The Growth of Animal Cells in Serum-Free Systems
IV.The Growth of Insect Cells in Serum-Free Systems
V.The Production of Viruses in Serum-Free Systems
VI.Problems Associated with the Use of Serum-Free Media and Its Current Position in Cell Culture
References
Chapter 16. A Rapid-Mixing Technique to Measure Transport in Suspended Animal Cells: Applications to Nucleoside Transport in Novikoff Rat Hepatoma Cells
I.Introduction
II.Apparatus and Procedures
III.Cells and Substrates
IV. Evaluation of Data
V.Conclusions
References
Chapter 17. Cloning of Normal and Transformed Cells on Plastic Films
I. Introduction
II.Standard Cloning Procedures
III.Cloning on Plastic Films
IV.Growth on Plastic Films
References
Chapter 18. A Simple Replica-Plating and Cloning Procedure for Mammalian Cells Using Nylon Cloth
I. Introduction
II.Replica Plating Using Nitex Nylon Cloth
III.Clone Isolation Using Nitex
IV.Conclusion
References
Note Added in Proof
Chapter 19. Human Minisegregant Cells
I.Introduction
II.Extrusion Subdivision and the Production of Minisegregant Cells
III.Isolation and Properties of HeLa Minisegregants
IV.The Fusion of Minisegregant and Other Cell Fragments and the Transfer of Genetic Information
V.Conclusions
VI.Appendix: The Assembly of a Pressure Chamber for Synchronizing Human Cells
References
Chapter 20. The Use of Tween-80-Permeabilized Mammalian Cells in Studies of Nucleic Acid Metabolism
I.Introduction
II. Methods
III.Results
IV. Concluding Remarks
References
Chapter 21. Nucleic Acid Synthesis in Permeabilized Eukaryotic Cells
I. Introduction
II. Materials
III. Procedure
IV. Characteristics of Nucleic Acid Synthesis
V. Summary
References
Chapter 22. Red Cell-Mediated Microinjection of Macromolecules into Mammalian Cells
I. Introduction
II. General Methods
III. Red Cell Loading
IV. Red Cell-Culture Cell Fusions
V. Concluding Remarks
References
Chapter 23. Selection of Somatic Cell Hybrids between HGPRT-and APRT - Cells
I. Introduction
II. Techniques
III. Results, Discussion, and Some Precautions
References
Chapter 24. Sequential Dissociation of the Exocrine Pancreas into Lobules, Acini, and Individual Cells
I. Introduction
II. Materials and Methods
III. Results and Discussion
References
Chapter 25. Bromodeoxyuridine Differential Chromatid Staining Technique: A New Approach to Examining Sister Chromatid Exchange and Cell Replication Kinetics
I. Introduction
II. Methodology
Ill. SCE Analysis for Examining Clastogenic Agents
IV. Assessment of the Spontaneity of SCE
V. Determination of Cellular Proliferation Kinetics
VI. Examination of DNA Replication Patterns
VII. Conclusions
References
Chapter 26. Preparation and Characterization of Mitochondria and Submitochondrial Particles of Rat Liver and Liver- Derived Tissues
I. Purpose
II. Advantages of Using Rat Liver Mitochondria for Studying Mitochondrial Biochemistry and Bioenergetics
III. Introductory Comments regarding Methods for Preparing Rat Liver Mitochondria
IV. Mitochondrial Isolation Media
V. Preparation of Rat Liver Mitochondria
VI. Additional Purification of Mitochondria Prepared by Differential Centrifugation
VII. Preparation of Rat Hepatoma Mitochondria
VIII. Preparation of Mitochondria from Regenerating Liver
IX. Introductory Comments regarding Submitochondrial Particle Preparations of Rat Liver
X. Preparation of Sonic Particles
XI. Preparation of Lubrol Particles
XII. Preparation of Mitochondrial Ghosts
XIII. Subfractionation of Mitochondria
XIV. Preparation of an Inner Membrane Fraction Deficient in ATPase Activity
XV. Preparation of an Inner Membrane Fraction Enriched in ATPase Activity ("Light" Membranes)
XVI. Introductory Comments regarding the Characterization of Mitochondria and Submitochondrial Particles for Intactness
References
Chapter 27. The Use of Complement-Mediated Cytolysis to Analyze Specific Cell Populations of Eukaryote Cells in Cultures
I. Introduction
II. Preparation of Reagents for Complement-Mediated Cytolysis
III. Cytotoxicity Assay
IV. Applications of Antibody-Complement Cytolysis
References
Chapter 28. Microelectrophoresis of Enzymes in Animal Cell Colonies
I. Introduction
II. Starch Gel Microelectrophoresis
III. Cellulose Acetate Microelectrophoresis
IV. Polyacrylamide Gel Microelectrophoresis
V. Microelectrophoresis of Single Cells
VI. Concluding Remarks
References
Subject Index
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