Epstein-Barr Virus Protocols

ISBN-13: 9781617371370 / Angielski / Miękka / 2010 / 438 str.

Joanna B. Wilson; Gerhard H. W. May

Opis

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."

Recenzje

"Timely and highly practical, Epstein-Barr Virus Protocols 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. Protocols 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 protocols 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

Spis treści

I. Genome and Transcript Analyses Epstein-Barr Virus: The B95-8 Strain Map Paul J. Farrell Analysis of Replication of oriP-Based Plasmids by Quantitative, Competitive PCR Ann L. Kirchmaier Genetic Analysis and Gene Expression with Mini-Epstein-Barr Virus Plasmids Ellen Kilger and Wolfgang Hammerschmidt Construction of cDNA Libraries for the Analysis of the Structure of Complementary Strand Transcripts (CSTs) Paul R. Smith. Analysis of the Expression and Function of the EBV-Encoded Small RNAs, the EBERs, in Heterologous Cells Kenneth G. Laing, Volker Matys, and Michael J. Clemens Visualizing EBV Expression Patterns by FISH Anna Szeles II. Viral Detection In Situ Detection of Epstein-Barr Virus DNA and Viral Gene Products Gerald Niedobitek and Hermann Herbst Phenotype Determination of Epstein-Barr Virus-Infected Cells in Tissue Sections Hermann Herbst and Gerald Niedobitek Detection of EBV Infection at the Single-Cell Level: Precise Quantitation of Virus-Infected Cells In Vivo Gregory J. Babcock, Emily M. Miyashita-Lin, and David A. Thorley-Lawson Detection and Discrimination of Latent and Replicative Herpesvirus Infection at the Single Cell Level In Vivo Lisa L. Decker, Gregory J. Babcock, and David A. Thorley-Lawson III. Culture Methods Virus Isolation Lindsey M. Hutt-Fletcher and Susan M. Turk Generation of Lymphoblastoid Cell Lines (LCLs) Teresa Frisan, Victor Levitsky, and Maria Masucci Cell Cycle Distribution of B-Lymphocytes and Cell Lines Alison J. Sinclair Introduction of Plasmid Vectors into Cells Via Electroporation Ann L. Kirchmaier Malignant Transformation and Immortalization Assays in Animal Cells Transfected with the BARF1 Gene Tadamasa Ooka Transient Gene Expression and MACS Enrichment Dieter Kube and Martina Vockerodt In Vitro Assays to Study Epithelial Cell Growth Christopher W. Dawson and Lawrence S. Young In Vitro Assays to Study Epithelial Cell Differentiation Christopher W. Dawson, and Lawrence S. Young Cell Sensitivity Assays: Quantitative Detection of Apoptotic Cells In Vitro Using the TUNEL Assay Neil A. Jones and Caroline Dive Qualitative Detection of Apoptotic Cells Assessed by DNA Fragmentation Amanda J. McIlwrath IV. Immune Assays Regression Assay Teresa Frisan, Victor Levitsky, and Maria Masucci Generation of Polyclonal EBV-Specific CTL Cultures and Clones Victor Levitsky, Teresa Frisan, and Maria Masucci Determination of Antigen and Fine Peptide Specificity of EBV-Specific CTLs Victor Levitsky, Teresa Frisan, and Maria Masucci Limiting Dilution Assay Teresa Frisan, Victor Levitsky, and Maria Masucci V. Viral Protein Detection Antibodies for Detecting EBV Latent Proteins Martin Rowe, and Anja Mehl Detection of EBV Latent Proteins by Western Blotting Martin Rowe and Matthew Jones Biosynthetic Radiolabeling of Virus Glycoproteins for Immunoprecipitation and Electrophoretic Analysis Lindsey M. Hutt-Fletcher and Susan M. Turk VI. Protein-Protein, Protein-DNA, and Protein-RNA Interactions The Yeast Two-Hybrid Assay to Identify Interacting Proteins Kenneth M. Izumi Identification of Transactivation, Repression, and Protein-Protein Interaction Domains Using GAL4-Fusion Proteins James J.-D. Hsieh and S. Diane Hayward Magnetic DNA Affinity Purification of a Cellular Transcription Factor Lothar J. Strobl and Ursula Zimber-Strobl Screening an Expression Library with a DNA Binding Site Qin Zhang and Eduardo A. Montalvo Analysis of DNA Binding Proteins by Mobility S

Blurb

The application of recombinant DNA technology to the analysis of Epstein-Barr virus (EBV) is rapidly developing sufficient insight into the virus-host interaction, so that its role in disease pathology is now often discernible and can increasingly be interdicted. In Epstein-Barr Virus Protocols, Joanna Wilson and Gerhard May have assembled a collection of the key molecular biology protocols used in the analysis of Epstein-Barr virus, along with a series of valuable immunology, cell biology, and transgenic mouse protocols. Described in step-by-step detail by experts who use them regularly, these readily reproducible techniques include methods for gene expression with mini-EBV plasmids, for expression analysis by FISH, for EBV detection and quantitation, and for cell proliferation and death assays. In addition, the authors provide information on EBV-based vectors, an up-to-date map of EBV, a comprehensive table of available latent protein antisera, and assays from in vitro to cell to organ to organism levels.
Timely and highly practical, Epstein-Barr Virus Protocols provides powerful tools for elucidating the life cycle of EBV and its host interactions, work that promises the emergence of major new treatments and cures for EBV-associated diseases, including several forms of human cancer.