Centrosomes and spindle pole bodies / edited by Robert E. Palazzo, Trisha N. Davis.

Format:

Book

Contributor:

Palazzo, Robert E.

Davis, Trisha, 1954-

Series:

Methods in cell biology ; 67.

Methods in cell biology ; 67

Language:

English

Subjects (All):

Centrosomes.

Spindle (Cell division).

Physical Description:

1 online resource (392 p.)

Place of Publication:

San Diego : Academic Press, c2001.

Language Note:

English

Summary:

Containing a comprehensive collection of convenient and quantitative methods for studying centrosomes, spindle pole bodies and related organelles, this text is a valuable resource for researchers and others interested in studying the role of these organelles in cell replication. Chapters outlining the role of these organelles in other cell functions are also included, and a wide variety of experimental systems for analyzing these organelles are presented. Detailed protocols for experiments are contained in each chapter for researchers to perform in their own labs. This volume outlines key meth

Contents:

Cover; Contents; Contributors; Preface; Chapter 1. Purification and Reconstitution of Drosophila y-Tubulin Complexes; I. Introduction; II. Isolation of γ -Tubulin-Containing Complexes from Drosophila Embryo Extracts; III. γTuSC Reconstitution in Sf9 Cells; IV. Functional Assays for Isolated γTuSC or γTuRC; V. Concluding Remarks; References; Chapter 2. Using Rapid Freeze and Freeze-Substitution for the Preparation of Yeast Cells for Electron Microscopy and Three-Dimensional Analysis; I. Introduction; II. How Rapid Freezing and Freeze-Substitution Work; III. Rapid Freezing Techniques

IV. Freeze-Substitution ProtocolsV. Embedding Protocols; VI. Sectioning and Imaging; VII. Immunolabeling for Electron Microscopy; VIII. Three-Dimensional Modeling of Spindle Pole Bodies and Other Organelles; IX. Conclusion; References; Chapter 3. Digital Fluorescence Microscopy of Cell Cytoplasts with and without the Centrosome; I. Introduction; II. Experimental Procedures; References; Chapter 4. Methods for the Study of Pericentrin in Centrosome Assembly and Function; I. Introduction; II. Pericentrin Localization to Centrosomes; III. Pericentrin Recruitment to Centrosomes

IV. Obtaining Enriched Fractions of PericentrinV. Artificial Elevation of Pericentrin Levels; VI. Pericentrin Staining to Detect Centrosome Defects in Tumor Tissues and Cell Lines; References; Chapter 5. Molecular Dissection of Yeast Spindle Pole Bodies by Two Hybrid, in Vitro Binding, and Co-purification; I. Introduction; II. The Two-Hybrid Approach; III. Immuno- and Affinity Precipitation of Spindle Pole Body Components; IV. In Vitro Binding of Spindle Pole Body Proteins; References; Chapter 6. Genetic Analysis of Yeast Spindle Pole Bodies; I. Introduction

II. Dosage-Dependent Suppressor ScreenIII. Extragenic Suppressor Screen; IV. Synthetic Lethal Screen; V. Synthetic Dosage Lethality; VI. Conclusions; References; Chapter 7. Methods for the Study of Centrosomes in Drosophila during Embryogenesis; I. Introduction; II. Materials; III. Methods; References; Chapter 8. Methods for Identification of Centrosome-Associated Proteins; I. Introduction; II. Preparation of Centrosomes and Anticentrosomal Antibodies; III. Molecular Cloning of cDNA Encoding Centrosome-Associated Proteins; IV. Evaluation of Clone Specificity

V. Identification of a Novel 135-kDa Centrosomal Protein, Cep135References; Chapter 9. Reconstitution of Centrosome Microtubule Nucleation in Drosophila; I. Introduction; II. Preparation of Extracts and Centrosomes from Drosophila Embryos; III. In Vitro Reconstitution of Microtubule Nucleation by Salt-Inactivated Centrosomes; IV. Conclusions; References; Chapter 10. Reconstitution of Centrosome Microtubule Nucleation in Spisula; I. Introduction; II. Preparations; III. Disassembly and Reassembly of Microtubule Nucleation Potential; IV. Fractionation of Extracts; V. Summary; References

Chapter 11. In Vitro Approaches for the Study of Microtubule Nucleation at the Fission Yeast Spindle Pole Body

Notes:

Description based upon print version of record.

Includes bibliographical references and index.

ISBN:

9786611012144

9780080925172

0080925170

9781281012142

1281012149

9780125439855

0125439857

9780080496627

0080496628

OCLC:

169914903