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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.
Contents of Previous Volumes
Chapter 1. Monoclonal Antibody Techniques Applied to Viruses
II. Immunization Protocol
III. Preparation of Cells
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
XII. Appendix: Preparation of Reagents
Chapter 2. Competition Radioimmunoassays for Characterization of Antibody Reactions to Viral Antigens
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
Chapter 3. Enzyme Immunosorbent Assays in Plant Virology
II. Principles of Enzyme Immunosorbent Assay
III. Preparation of Sample
IV. Production of Antiserum
V. Choice of Enzyme and Preparation of Conjugate
VII. Choice of Solid Phase
VIII. Types of ELISA Procedure
IX. Evaluation of Reagents and Analysis of Results
XI. Appendix: Buffer Formulations
Chapter 4. Electron Microscopy for the Identification of Plant Viruses in in Vitro Preparations
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
Chapter 5. Cloning and Expression of Viral Antigens in Escherichia coli and Other Microorganisms
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
Chapter 6. Spot Hybridization for Detection of Viroids and Viruses
II. PSTV Diagnosis by Nucleic Acid Spot Hybridization
III. Application to Other Viroid Diseases
IV. Application to Viral Diseases
V. Supplementary Methodologies
Chapter 7. Detection of Viral Nucleic Acids by in Situ Hybridization
II. Materials and Methods
Chapter 8. Exploring the Gene Organization of Baculoviruses
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
Chapter 9. Detection and Characterization of Subgenomic RNAs in Plant Viruses
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
- No. of pages:
- © Academic Press 1984
- 11th October 1984
- Academic Press
- eBook ISBN:
Professor Karl Maramorosch works at Rutgers University, New Brunswick, NJ, USA.
Rutgers University, New Brunswick, NJ, USA
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