Epstein-Barr Virus Protocols

About this book

Joanna Wilson and Gerhard May have assembled a collection of the key molecular biology protocols used in the analysis of Epstein-Barr virus (EBV), along with a series of valuable immunology, cell biology, and transgenic mouse protocols. 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, there are 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.

Contents

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 Shift Assay Yi-Chun James Wang and Eduardo A. Montalvo Analysis of RNA-Protein Interactions of the EBV-Encoded Small RNAs, the EBERs: In Vitro Assays Kenneth G. Laing, Androulla Elia, Ian W. Jeffrey, and Michael J. Clemens VII. Protein Activity Assays Chimeric and Mutated Variants of LMP1: A Helpful Tool to Analyze the Structure-Function Relationship of a Pseudoreceptor Olivier Gires, Marius Ueffing, and Wolfgang Hammerschmidt Assaying the Activity of Kinases Regulated by LMP1 Arnd Kieser In Vitro Assays for the Detection of Protein Tyrosine Phosphorylation and Protein Tyrosine Kinase Activities Sara Fruehling and Richard Longnecker VIII. Tissue and In Vivo Protocols Analysis of Apoptosis in Tissue Sections Vicki Save, Philip J. Coates, and Peter A. Hall Considerations in Generating Transgenic Mice: DNA, RNA, and Protein Extractions from Tissues - Rapid and Effective Blotting Joanna B. Wilson and Mark E. Drotar In Vivo Assay of Cellular Proliferation John Curran Topical Chemical Carcinogen Treatment in Mice John Curran Separation of Epidermal Tissue from Underlying Dermis and Primary Keratinocyte Culture Jennifer Macdiarmid and Joanna B. Wilson Selection and Enrichment of B Cells from Lymphoid Tissues Penelope Tsimbouri, Marie Anne O'Donnell, and Joanna B. Wilson Detection of Immunoglobulin Gene Rearrangements Robert G. Caldwell and Richard Longnecker

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