Maramorosch / Koprowski Methods in Virology

Volume V
1. Auflage 2014
ISBN: 978-1-4832-6223-9
Verlag: Elsevier Science & Techn.
Format: PDF
Kopierschutz: 1 - PDF Watermark

E-Book, Englisch, 548 Seiten, Web PDF

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.

Autoren/Hrsg.

Maramorosch, Karl

Koprowski, Hilary

Inhaltsverzeichnis

1;Front Cover;1
2;Methods in Virology;4
3;Copyright Page;5
4;Table of Contents;6
5;List of Contributors;12
6;Preface;14
7;Contents of Other Volumes;16
8;Chapter 1. Fusion of Cells for Virus Studies and Production of Cell Hybrids;22
8.1;I. Introduction;22
8.2;II. Methods;26
8.3;III. Cell Fusion in Virology;40
8.4;IV. Selection of Hybrid Cell Lines;44
8.5;V. Uses of Heterokaryons and Hybrid Cell Lines;51
8.6;References;52
9;Chapter 2. New Approaches to Ultracentrifugation;54
9.1;I. Introduction;55
9.2;II. A Solution: New Approaches;56
9.3;III. Results;71
9.4;IV. Discussion and Summary;95
9.5;Acknowledgments;97
9.6;References;97
10;Chapter 3. Small Circular Viral DNA: Preparation and Analysis;100
10.1;I. Introduction;100
10.2;II. Virus Cultivation, Concentration, and Purification;103
10.3;III. DNA Extraction and Purification;112
10.4;IV. Physical Analysis of DNA Components;124
10.5;V. Biological Analysis of DNA Components;130
10.6;Appendix;141
10.7;Acknowledgments;142
10.8;References;143
11;Chapter 4. Polyacrylamide Gel Electrophoresis of Viral RNA;146
11.1;I. Introduction;147
11.2;II. Apparatus;148
11.3;III. Procedures for Preparation and Running of Gels;151
11.4;IV. Analysis of the Distribution of RNA on Polyacrylamide Gels after Electrophoresis;166
11.5;V. Elution of RNO from Analytical Gel Fractions;180
11.6;VI. Large-Scale Preparative Techniques;181
11.7;VII. Applications to Virological Problems and the Analysis of Results;185
11.8;VIII. Conclusion;196
11.9;Acknowledgments;196
11.10;References;196
12;Chapter 5. Polyacrylamide Gel Electrophoresis of Viral Proteins;200
12.1;I. Introduction;201
12.2;II. Procedures for Acrylamide Gel Electrophoresis;202
12.3;III. Interpretation of Results;246
12.4;IV. Discussion;251
12.5;V. Appendix: Common Complaints, Troubleshooting Suggestions, and Tips;261
12.6;References;265
13;Chapter 6. DNA—RNA and DNA—DNA Hybridization in Virus Research;268
13.1;I. Introduction;269
13.2;II. Materials;271
13.3;III. DNA—RNA Hybridization Procedure;273
13.4;IV. Characteristics of the DNA—RNA Hybridization Reaction;275
13.5;V. Thermal Elution of RNA from RNA—DNA Hybrids;279
13.6;VI. Hybridization and Elution Procedures that Minimize RNA Degradation;282
13.7;VII. DNA—DNA Hybridization Procedure;283
13.8;VIII. Alternative Methods of Hybridization;285
13.9;Acknowledgment;289
13.10;References;289
14;Chapter 7. Techniques of RNA—DNA Hybridizationin Solution for the Study of Viral Transcription;292
14.1;I. Introduction;292
14.2;II. Basic Equipment;294
14.3;III. Preparation of Nucleic Acids;295
14.4;IV. RNA—DNA Hybridization in Solution (Liquid Hybridization);299
14.5;V. Segmental Transcription Studies by RNA—DNA Hybridization in Solution;302
14.6;References;312
15;Chapter 8. Methods for the Study of mRNA Synthesis in Bacteriophage-Infected E. coli;314
15.1;I. General Introduction;314
15.2;II. Bacteriophage–Host System;315
15.3;III. Isolation Methods;320
15.4;IV. Analytical Methods;334
15.5;Acknowledgments;358
15.6;References;358
16;Chapter 9. Reactions of Viruses in Agar Gel;368
16.1;I. Introduction;368
16.2;II. Reactions between Viral Antigens and Antibodies;369
16.3;III. Reactions between Viruses and Erythrocyte Stromata;388
16.4;IV. Electron Microscopic Examination of Reaction Lines;389
16.5;Acknowledgments;391
16.6;References;391
17;Chapter 10. Methods for the Study of Virus–Antibody Complexes;396
17.1;I. General Introduction;396
17.2;II. Methods of Reactivating Neutralized Virus;399
17.3;III. Characterization of the Virus–Antibody Complex;407
17.4;IV. Methods of Separating the Components of a Virus–Antibody Reaction Mixture;415
17.5;References;417
18;Chapter 11. Techniques of Ferritin- Tagged Antibodies;420
18.1;I. Introduction;420
18.2;II. Purification of Ferritin;421
18.3;III. Preparation of Antibody Globulin Fraction from Serum;425
18.4;IV. Conjugation of Ferritin and Antibody;427
18.5;V. Evaluating the Conjugate;431
18.6;VI. Methods of Direct Ferritin-Tagging;436
18.7;VII. Methods of Indirect Ferritin-Tagging;438
18.8;VIII. Controls in Ferritin-Tagging Experiments;439
18.9;IX. Hybrid Antibody Method for Ferritin Localization of Antigen;440
18.10;X. Application of Hybrid Antibody;441
18.11;XI. Other Methods of Visualizing Immune Globulins;441
18.12;XII. Conclusions;441
18.13;References;442
19;Chapter 12. The Immunoperoxidase Technique: Localization of Viral Antigens in Cells;444
19.1;I. Introduction;444
19.2;II. The Principles of the Immunoperoxidase Technique;446
19.3;III. Properties of Horseradish Peroxidase;447
19.4;IV. Histochemical Detection of Peroxidase Activity;447
19.5;V. Peroxidase-Antibody Labeling Techniques;450
19.6;VI. Procedure for Detection of Viral Antigens in Cells by the Immunoperoxidase Technique;452
19.7;VII. Interpretation of the Results Obtained for the Localization of Viral Antigens in Cells by the Immunoperoxidase Technique;455
19.8;Acknowledgments;463
19.9;References;463
20;Chapter 13. Leaf Dip Serology;466
20.1;I. Introduction;466
20.2;II. Procedure;467
20.3;III. Results and Discussion;468
20.4;References;471
21;Chapter 14. Procedures to Increase Virus Yield from Infected Plants;472
21.1;I. Introduction;473
21.2;II. Sequence of Events in Virus Infection;473
21.3;III. Criteria for Virus Yield;475
21.4;IV. Factors Affecting Virus Yield;479
21.5;V. Summary;495
21.6;References;495
22;Chapter 15. Freeze-Etching Technique for the Study of Virus Ultrastructure;502
22.1;I. Introduction;502
22.2;II. The Freeze-Etching Technique;503
22.3;III. Results up to the Present;509
22.4;IV. Conclusion and Prospects;514
22.5;Acknowledgments;515
22.6;References;515
23;Author Index;518
24;Subject Index;537