Epstein-Barr Virus Protocols -

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Résumé :
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.

Sommaire:
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 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 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 Limiting Dilution Assay 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 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