Methods in Virology (eBook)

Volume V

Hilary Koprowski, Karl Maramorosch (Herausgeber)

eBook Download: PDF

2014 | 1. Auflage
548 Seiten
Elsevier Science (Verlag)
978-1-4832-6223-9 (ISBN)

Infos

Methods in Virology, Volume V focuses on the methods used in virology, including hybridization, gel electrophoresis, freeze-etching technique, and ultracentrifugation. The selection first offers information on the fusion of cells for virus studies and production of cell hybrids; approaches to ultracentrifugation; and polyacrylamide gel electrophoresis of viral RNA. Discussions focus on applications to virological problems and analysis of results; analysis of the distribution of RNA on polyacrylamide gels after electrophoresis; and biological analysis of DNA components. The book then examines the polyacrylamide gel electrophoresis of viral proteins, DNA-RNA and DNA-DNA hybridization in virus research, and techniques of RNA-DNA hybridization in solution for the study of viral transcription. Topics include preparation of nucleic acids, hybridization and elution procedures that minimize RNA degradation, and procedures for acrylamide gel electrophoresis. The text takes a look at freeze-etching technique for the study of virus ultrastructure; procedures to increase virus yield from infected plants; and the immunoperoxidase technique. Concerns include principles of the immunoperoxidase technique, histochemical detection of peroxidase activity, sequence of events in virus infection, and factors affecting virus yield. The selection is a valuable source of data for researchers interested in the methods employed in virology.

Inhalt

Front Cover 1
Methods in Virology 4
Copyright Page 5
Table of Contents 6
List of Contributors 12
Preface 14
Contents of Other Volumes 16
Chapter 1. Fusion of Cells for Virus Studies and Production of Cell Hybrids 22
I. Introduction 22
II. Methods 26
III. Cell Fusion in Virology 40
IV. Selection of Hybrid Cell Lines 44
V. Uses of Heterokaryons and Hybrid Cell Lines 51
References 52
Chapter 2. New Approaches to Ultracentrifugation 54
I. Introduction 55
II. A Solution: New Approaches 56
III. Results 71
IV. Discussion and Summary 95
Acknowledgments 97
References 97
Chapter 3. Small Circular Viral DNA: Preparation and Analysis 100
I. Introduction 100
II. Virus Cultivation, Concentration, and Purification 103
III. DNA Extraction and Purification 112
IV. Physical Analysis of DNA Components 124
V. Biological Analysis of DNA Components 130
Appendix 141
Acknowledgments 142
References 143
Chapter 4. Polyacrylamide Gel Electrophoresis of Viral RNA 146
I. Introduction 147
II. Apparatus 148
III. Procedures for Preparation and Running of Gels 151
IV. Analysis of the Distribution of RNA on Polyacrylamide Gels after Electrophoresis 166
V. Elution of RNO from Analytical Gel Fractions 180
VI. Large-Scale Preparative Techniques 181
VII. Applications to Virological Problems and the Analysis of Results 185
VIII. Conclusion 196
Acknowledgments 196
References 196
Chapter 5. Polyacrylamide Gel Electrophoresis of Viral Proteins 200
I. Introduction 201
II. Procedures for Acrylamide Gel Electrophoresis 202
III. Interpretation of Results 246
IV. Discussion 251
V. Appendix: Common Complaints, Troubleshooting Suggestions, and Tips 261
References 265
Chapter 6. DNA—RNA and DNA—DNA Hybridization in Virus Research 268
I. Introduction 269
II. Materials 271
III. DNA—RNA Hybridization Procedure 273
IV. Characteristics of the DNA—RNA Hybridization Reaction 275
V. Thermal Elution of RNA from RNA—DNA Hybrids 279
VI. Hybridization and Elution Procedures that Minimize RNA Degradation 282
VII. DNA—DNA Hybridization Procedure 283
VIII. Alternative Methods of Hybridization 285
Acknowledgment 289
References 289
Chapter 7. Techniques of RNA—DNA Hybridizationin Solution for the Study of Viral Transcription 292
I. Introduction 292
II. Basic Equipment 294
III. Preparation of Nucleic Acids 295
IV. RNA—DNA Hybridization in Solution (Liquid Hybridization) 299
V. Segmental Transcription Studies by RNA—DNA Hybridization in Solution 302
References 312
Chapter 8. Methods for the Study of mRNA Synthesis in Bacteriophage-Infected E. coli 314
I. General Introduction 314
II. Bacteriophage–Host System 315
III. Isolation Methods 320
IV. Analytical Methods 334
Acknowledgments 358
References 358
Chapter 9. Reactions of Viruses in Agar Gel 368
I. Introduction 368
II. Reactions between Viral Antigens and Antibodies 369
III. Reactions between Viruses and Erythrocyte Stromata 388
IV. Electron Microscopic Examination of Reaction Lines 389
Acknowledgments 391
References 391
Chapter 10. Methods for the Study of Virus–Antibody Complexes 396
I. General Introduction 396
II. Methods of Reactivating Neutralized Virus 399
III. Characterization of the Virus–Antibody Complex 407
IV. Methods of Separating the Components of a Virus–Antibody Reaction Mixture 415
References 417
Chapter 11. Techniques of Ferritin- Tagged Antibodies 420
I. Introduction 420
II. Purification of Ferritin 421
III. Preparation of Antibody Globulin Fraction from Serum 425
IV. Conjugation of Ferritin and Antibody 427
V. Evaluating the Conjugate 431
VI. Methods of Direct Ferritin-Tagging 436
VII. Methods of Indirect Ferritin-Tagging 438
VIII. Controls in Ferritin-Tagging Experiments 439
IX. Hybrid Antibody Method for Ferritin Localization of Antigen 440
X. Application of Hybrid Antibody 441
XI. Other Methods of Visualizing Immune Globulins 441
XII. Conclusions 441
References 442
Chapter 12. The Immunoperoxidase Technique: Localization of Viral Antigens in Cells 444
I. Introduction 444
II. The Principles of the Immunoperoxidase Technique 446
III. Properties of Horseradish Peroxidase 447
IV. Histochemical Detection of Peroxidase Activity 447
V. Peroxidase-Antibody Labeling Techniques 450
VI. Procedure for Detection of Viral Antigens in Cells by the Immunoperoxidase Technique 452
VII. Interpretation of the Results Obtained for the Localization of Viral Antigens in Cells by the Immunoperoxidase Technique 455
Acknowledgments 463
References 463
Chapter 13. Leaf Dip Serology 466
I. Introduction 466
II. Procedure 467
III. Results and Discussion 468
References 471
Chapter 14. Procedures to Increase Virus Yield from Infected Plants 472
I. Introduction 473
II. Sequence of Events in Virus Infection 473
III. Criteria for Virus Yield 475
IV. Factors Affecting Virus Yield 479
V. Summary 495
References 495
Chapter 15. Freeze-Etching Technique for the Study of Virus Ultrastructure 502
I. Introduction 502
II. The Freeze-Etching Technique 503
III. Results up to the Present 509
IV. Conclusion and Prospects 514
Acknowledgments 515
References 515
Author Index 518
Subject Index 537

Produkt-Details

• Erscheint lt. Verlag: 28.6.2014
• Sprache: englisch
• Themenwelt: Medizin / Pharmazie ► Medizinische Fachgebiete
• Themenwelt: Studium ► Querschnittsbereiche ► Infektiologie / Immunologie
• Themenwelt: Naturwissenschaften ► Biologie ► Evolution
• Themenwelt: Technik
• ISBN-10: 1-4832-6223-5 / 1483262235
• ISBN-13: 978-1-4832-6223-9 / 9781483262239