DNA Viruses Methods and Protocols

A compendium of readily reproducible and novel methods to manipulate DNA viruses and characterize their varied biological properties. The authors emphasize techniques for viral detection and genetics, but also include methods for structure determination, gene expression, replication, pathogenesis, complex cellular models, recombinant genetics, and computational/systems approaches. Wide-ranging and highly practical, DNA Viruses: Methods and Protocols will stimulate new directions in virology research with its novel strategies for engineering viral vectors in gene therapy, and its advanced approaches for detecting viruses in human disease.

Viral Detection.- Viral Detection.- Quantitative Detection of Epstein-Barr Virus DNA in Clinical Specimens by Rapid Real-Time PCR Targeting a Highly Conserved Region of EBNA-1.- Profiling of Epstein-Barr Virus Latent RNA Expression in Clinical Specimens by Gene-Specific Multiprimed cDNA Synthesis and PCR.- Quantitative Detection of Viral Gene Expression in Populations of Epstein-Barr Virus-Infected Cells In Vivo.- Detection and Quantification of the Rare Latently Infected Cell Undergoing Herpes Simplex Virus Transcriptional Activation in the Nervous System In Vivo.- Reporter Cell Lines for the Detection of Herpes Simplex Viruses.- Virus Structure And Imaging.- Unraveling the Architecture of Viruses by High-Resolution Atomic Force Microscopy.- Studying the Structure of Large Viruses With Multiresolution Imaging.- Herpes Simplex Virus-Cell Interactions Studied by Low-Fading Contrasted Immunofluorescence.- Herpes Simplex Virus-Cell Interactions Studied by Immunogold Cryosection Electron Microscopy.- FTIR Microscopy Detection of Cells Infected With Viruses.- Virus Entry.- The JC Virus-Like Particle Overlay Assay.- Analysis of Fusion Using a Virus-Free Cell Fusion Assay.- Pseudovirions as Specific Tools for Investigation of Virus Interactions With Cells.- Gene Expression.- Simultaneous In Situ Detection of RNA, DNA, and Protein Using Tyramide-Coupled Immunofluorescence.- Identification and Characterization of Herpesviral Immediate-Early Genes.- Replication And Genome Maintenance.- Methods for Measuring the Replication and Segregation of Epstein-Barr Virus-Based Plasmids.- DNA Affinity Purification of Epstein-Barr Virus OriP-Binding Proteins.- Pathogenesis.- Pre-B-Cell Colony Formation Assay.- Luciferase Real-Time Bioluminescence Imaging for the Study of Viral Pathogenesis.-Complex Cell Systems.- Culturing Primary and Transformed Neuronal Cells for Studying Pseudorabies Virus Infection.- Human Papillomavirus Type 31 Life Cycle Methods for Study Using Tissue Culture Models.- Recombinant Genetics.- Molecular Genetics of Herpesviruses A Recombinant Technology Approach.- Molecular Genetics of DNA Viruses Recombinant Virus Technology.- Genetic Analysis of Cytomegalovirus by Shuttle Mutagenesis.- Construction of a Gene Inactivation Library for Bovine herpesvirus 1 Using Infectious Clone Technology.- Selective Silencing of Viral Gene E6 and E7 Expression in HPV-Positive Human Cervical Carcinoma Cells Using Small Interfering RNAs.- Computation/Systems Biology Of Viruses.- Design of a Herpes Simplex Virus Type 2 Long Oligonucleotide-Based Microarray.- Real-Time Quantitative PCR Analysis of Viral Transcription.- Rapid Screening of Chemical Inhibitors That Block Processive DNA Synthesis of Herpesviruses.