Epstein-Barr Virus Protocols / Edition 1

by Joanna B. Wilson, Gerhard H. W. May

Overview

The discovery of Epstein-Barr virus (EBV) by Epstein, Achong, and Barr, reported in 1964 (Lancet 1:702–703), was stimulated by Denis Burkitt’s rec- nition of a novel African childhood lymphoma and his postulation that an infectious agent was involved in the tumor’s etiology (Nature194:232–234, 1962). Since then, molecular and cellular biological and computational technologies have progressed by leaps and bounds. The advent of recombinant DNA technology opened the possibilities of genetic research more than most would have realized. Not only have the molecular tools permitted the analyses of viral mechanisms, but, importantly, they have formed the basis for discerning viral presence and, subsequently, viral involvement in an increasing number of diseases. Though in every field of science the search for further knowledge is likely to be a limitless phenomenon, the distinct goal in EBV research, namely, to gain sufficient insight into the viral–host interaction to be able to intercept the pathogenic process, is beginning to be realized. Epstein-Barr virus research has effectively entered the postgenomic era that began with the sequencing of the first strains, cloned in the mid to late 1980s.

Product Details

ISBN-13:	9781617371370
Publisher:	Springer-Verlag New York, LLC
Publication date:	11/10/2010
Series:	Methods in Molecular Biology , #174
Edition description:	Softcover reprint of hardcover 1st ed. 2001
Pages:	438
Product dimensions:	6.60(w) x 10.00(h) x 1.20(d)

About the Author

Wilson, Joanna B.; May, Gerhard H. W.

Table of Contents

Genome and Transcript Analyses.- Epstein-Barr Virus.- Analysis of Replication of oriP-Based Plasmids by Quantitative, Competitive PCR.- Genetic Analysis and Gene Expression with Mini-Epstein-Barr Virus Plasmids.- Construction of cDNA Libraries for the Analysis of the Structure of Complementary Strand Transcripts (CSTs).- Analysis of the Expression and Function of the EBV-Encoded Small RNAs, the EBERs, in Heterologous Cells.- Visualizing EBV Expression Patterns by FISH.- Viral Detection.- In Situ Detection of Epstein-Barr Virus DNA and Viral Gene Products.- Phenotype Determination of Epstein-Barr Virus-Infected Cells in Tissue Sections.- Detection of EBV Infection at the Single-Cell Level.- Detection and Discrimination of Latent and Replicative Herpesvirus Infection at the Single Cell Level In Vivo.- Culture Methods.- Virus Isolation.- Generation of Lymphoblastoid Cell Lines (LCLs).- Cell Cycle Distribution of B-Lymphocytes and Cell Lines.- of Plasmid Vectors into Cells Via Electroporation.- Malignant Transformation and Immortalization Assays in Animal Cells Transfected with the BARF1 Gene.- Transient Gene Expression and MACS Enrichment.- In Vitro Assays to Study Epithelial Cell Growth.- In Vitro Assays to Study Epithelial Cell Differentiation.- Cell Sensitivity Assays.- Qualitative Detection of Apoptotic Cells Assessed by DNA Fragmentation.- Immune Assays.- Regression Assay.- Generation of Polyclonal EBV-Specific CTL Cultures and Clones.- Determination of Antigen and Fine Peptide Specificity of EBV-Specific CTLs.- Limiting Dilution Assay.- Viral Protein Detection.- Antibodies for Detecting EBV Latent Proteins.- Detection of EBV Latent Proteins by Western Blotting.- Biosynthetic Radiolabeling of Virus Glycoproteins for Immunoprecipitation and Electrophoretic Analysis.- Protein-Protein, Protein-DNA, and Protein-RNA Interactions.- The Yeast Two-Hybrid Assay to Identify Interacting Proteins.- Identification of Transactivation, Repression, and Protein-Protein Interaction Domains Using GAL4-Fusion Proteins.- Magnetic DNA Affinity Purification of a Cellular Transcription Factor.- Screening an Expression Library with a DNA Binding Site.- Analysis of DNA Binding Proteins by Mobility Shift Assay.- Analysis of RNA-Protein Interactions of the EBV-Encoded Small RNAs, the EBERs.- Protein Activity Assays.- Chimeric and Mutated Variants of LMP1.- Assaying the Activity of Kinases Regulated by LMP1.- In Vitro Assays for the Detection of Protein Tyrosine Phosphorylation and Protein Tyrosine Kinase Activities.- Tissue and In Vivo Prools.- Analysis of Apoptosis in Tissue Sections.- Considerations in Generating Transgenic Mice.- In Vivo Assay of Cellular Proliferation.- Topical Chemical Carcinogen Treatment in Mice.- Separation of Epidermal Tissue from Underlying Dermis and Primary Keratinocyte Culture.- Selection and Enrichment of B Cells from Lymphoid Tissues.- Detection of Immunoglobulin Gene Rearrangements.

Editorial Reviews

"Timely and highly practical, Epstein-Barr Virus Prools provides powerful tools for elucidating life cycle of EBV and its host interactions, work that promises the emergence of major new treatments and cures of EVB-associated diseases, including several forms of human cancer." - Anticancer Research

"Overall it is an excellent resource aimed at postgraduate students and young researchers. Prools are extensively referenced to the primary literature at the end of each chapter." -Today's Life Science

"This book provides a well-written and concise account of many of the prools currently in use for the study of the Epstein-Barr virus and its related diseases. It is comprehensive..." "Both volumes are laid out attractively...and contain much valuable information on the pharmacological aspects of neurotoxicology, as well as state-of-the-art methodology." -Microbiology Today

"Leading-edge and forward-looking, Tumor Suppressor Genes in Human Cancer illuminates what is currently known of tumor suppressor genes and their regulation, work that is already beginning to revolutionize, and treatment design." -Tumori

"This book is highly recommended for collections serving researchers in the area of molecular biology." -E-Streams

From the Publisher

3 Stars from Doody

International specialists in biological methods, cell biology, molecular biology, virology, biochemistry and immunology present information in eight major areas: genome and transcript analyses; viral detection; culture methods; immune assays; viral protein detection; protein-protein, protein-DNA and protein-RNA interactions; protein activity assays; and tissue and in vivo protocols. Included are methods for gene expression with mini-EBV plasmids, for expression analysis by FISH, for EBV detection and quantification, and for cell proliferation and death assays. Also included is information on EBV- based vectors, an up-to-date map of EBV, a table of available latent protein antisera, and assays from in vitro to cell to organ to organism levels. Techniques are presented in step-by-step detail to facilitate easy replication. Included are many methods suitable to related viruses (HHV8) and nonviral systems. Annotation c. Book News, Inc., Portland, OR (booknews.com)

Booknews

Reviewer: Rosemarie A Pereira, PhD (The University of Texas Medical Branch)
Description: This book in the Methods in Molecular Biology series provides an overview of principal methods of use to the Epstein-Barr virus (EBV) researcher. Written by experts in the field, the chapters review protocols to investigate the molecular biology and pathogenesis of, and immune response to, the virus.
Purpose: Recent advances in molecular biology and immunology have generated sophisticated methodology to address viral-host interactions more effectively. This timely and needed book aims to provide EBV researchers with principal, state-of-the-art methods to enhance understanding of pathogenesis and aid researchers in their ultimate quest for mechanisms to intercept viruses.
Audience: This book would be useful to researchers and students interested in EBV. Many of the protocols described here are adaptable to areas of research unrelated to EBV. The editors are highly skilled in the areas of molecular biology and immunology and experts in diverse fields of EBV research have written the chapters.
Features: The eight sections of the book cover a comprehensive range of methods, the majority of which are molecular biological. Much-needed descriptions of culture techniques and in situ analyses are also included. The final section describing in vivo protocols is an excellent conclusion to a well-organized book. Most of the references are recent. There is a reasonable amount of overlap in some chapters, which in my opinion is unavoidable.
Assessment: This book is well written and comprehensive. It provides a great deal of information to the EBV researcher and overviews current protocols in the field effectively. It should be a good reference source for researchers and students of herpes virology.

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