Methods in Virology

Volume VII

1st Edition - October 11, 1984
Imprint: Academic Press
Editors: Karl Maramorosch, Hilary Koprowski
Language: English
Paperback ISBN:
9 7 8 - 1 - 4 8 3 2 - 4 3 1 8 - 4
eBook ISBN:
9 7 8 - 1 - 4 8 3 2 - 6 9 5 0 - 4

Methods in Virology, Volume VII focuses on the methods used in virology, including radioimmunoassays, microscopy, hybridization, and mutagenesis. The selection first elaborates on… Read more

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Methods in Virology, Volume VII focuses on the methods used in virology, including radioimmunoassays, microscopy, hybridization, and mutagenesis. The selection first elaborates on monoclonal antibody techniques applied to viruses; competition radioimmunoassays for characterization of antibody reactions to viral antigens; and enzyme immunosorbent assays in plant virology. Discussions focus on the principles of enzyme immunosorbent assay, choice of enzyme and preparation of conjugate, determination of immunoglobulin class, and maintenance and specificity testing of hybridomas. The text then elaborates on electron microscopy for the identification of plant viruses in in vitro preparations and cloning and expression of viral antigens in Escherichia coli and other microorganisms, including influenza virus, expression of foreign coding sequences in Escherichia coli, hepatitis B virus, electron microscope, immunoelectron microscopy, and imaging of nucleic acids. The manuscript takes a look at the detection and characterization of subgenomic RNA in plant viruses; exploring the gene organization of baculoviruses; and spot hybridization for detection of viroids and viruses. Topics include application to viral diseases, mapping mutuations of baculoviruses, transcriptional mapping of baculovirus genomes, and genetic mapping by blot hybridization. The selection is a valuable source of information for researchers interested in the methods employed in virology.

Contributors

Preface

Contents of Previous Volumes

Chapter 1. Monoclonal Antibody Techniques Applied to Viruses

I. Introduction

II. Immunization Protocol

III. Preparation of Cells

IV. Hybridization

V. Maintenance and Specificity Testing of Hybridomas

VI. Cloning of Hybridomas

VII. Determination of Immunoglobulin Class

VIII. Preparation of Ascites Fluid

IX. Concentration and Purification of Immunoglobulins

X. Characterization of Viral Antigens Using Monoclonal Antibodies

XI. Prospectus

XII. Appendix: Preparation of Reagents

References

Chapter 2. Competition Radioimmunoassays for Characterization of Antibody Reactions to Viral Antigens

I. Introduction

II. Preparation and Characterization of Reagents

III. Basic Methodologies

IV. Applicability to Identification of Antibody Populations

V. Applicability to Characterization of Viral Antigens

VI. Biological Tracking of Viral Proteins

VII. Concluding Comments

References

Chapter 3. Enzyme Immunosorbent Assays in Plant Virology

I. Introduction

II. Principles of Enzyme Immunosorbent Assay

III. Preparation of Sample

IV. Production of Antiserum

V. Choice of Enzyme and Preparation of Conjugate

VI. Substrates

VII. Choice of Solid Phase

VIII. Types of ELISA Procedure

IX. Evaluation of Reagents and Analysis of Results

X. Conclusions

XI. Appendix: Buffer Formulations

References

Chapter 4. Electron Microscopy for the Identification of Plant Viruses in in Vitro Preparations

I. Introduction

II. The Electron Microscope

III. Support Films

IV. Calibration of Magnification

V. Negative Staining

VI. Metal Shadowing

VII. Immunoelectron Microscopy

VIII. Imaging of Nucleic Acids

References

Chapter 5. Cloning and Expression of Viral Antigens in Escherichia coli and Other Microorganisms

I. Introduction

II. Expression of Foreign Coding Sequences in Escherichia coli

III. Hepatitis B Virus

IV. Foot-and-Mouth Disease Virus

V. Poliovirus: Purification of Polio RNA and Cloning of Poliovirus cDNA

VI. Influenza Virus

VII. Rabies Virus

VIII. Expression of Viral Antigens in Bacillus subtilis

IX. Expression of Viral Antigens in Saccharomyces cerevisiae

X. Conclusion

References

Chapter 6. Spot Hybridization for Detection of Viroids and Viruses

I. Introduction

II. PSTV Diagnosis by Nucleic Acid Spot Hybridization

III. Application to Other Viroid Diseases

IV. Application to Viral Diseases

V. Supplementary Methodologies

References

Chapter 7. Detection of Viral Nucleic Acids by in Situ Hybridization

I. Introduction

II. Materials and Methods

III. Applications

References

Chapter 8. Exploring the Gene Organization of Baculoviruses

I. Introduction

II. Establishing the Physical Map of a Baculovirus

III. Mapping Mutations of Baculoviruses

IV. Correlating Proteins with the Physical Map

V. Transcriptional Mapping of Baculoviruses Genomes

VI. DNA Sequence Homology to Facilitate Mapping Baculoviruses Other than AcNPV

VII. Site-Directed Mutagenesis and Allelic Replacement

VIII. Summary

IX. Addendum

References

Chapter 9. Detection and Characterization of Subgenomic RNAs in Plant Viruses

I. Introduction

II. Detection and Characterization of Encapsidated Viral Subgenomic RNAs

III. Detection and Characterization of Subgenomic Viral RNAs in Extracts of Infected Tissues

IV. Genetic Mapping by Blot Hybridization

V. Concluding Remarks

References

Index

Life Sciences

Physical Sciences & Engineering

Social Sciences & Humanities

Health

Methods in Virology

Volume VII

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Karl Maramorosch

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