Methods in Virology

ContributorsPrefaceContents of Previous VolumesChapter 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 ReferencesChapter 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 ReferencesChapter 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 ReferencesChapter 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 ReferencesChapter 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 ReferencesChapter 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 ReferencesChapter 7. Detection of Viral Nucleic Acids by in Situ Hybridization I. Introduction II. Materials and Methods III. Applications ReferencesChapter 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 ReferencesChapter 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 ReferencesIndex