Methods in Virology

Methods in Virology

Volume VII

1st Edition - October 11, 1984

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  • Editors: Karl Maramorosch, Hilary Koprowski
  • eBook ISBN: 9781483269504

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Description

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.

Table of Contents


  • 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

Product details

  • No. of pages: 348
  • Language: English
  • Copyright: © Academic Press 1984
  • Published: October 11, 1984
  • Imprint: Academic Press
  • eBook ISBN: 9781483269504

About the Editors

Karl Maramorosch

Professor Karl Maramorosch works at Rutgers University, New Brunswick, NJ, USA.

Affiliations and Expertise

Professor, Rutgers University, New Brunswick, NJ, USA

Hilary Koprowski

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