Recombinant vaccinia viruses have become important in recent years, both as protein expression vectors, and-in the age of bioterrorism-as powerful tools for developing new smallpox vaccines, diagnostics, and understanding of poxvirus pathogenesis. In Vaccinia Virus and Poxvirology: Methods and Protocols, internationally recognized experts and leaders in poxvirology describe in step-by-step detail the successful techniques they have perfected to study and work with vaccinia virus and other poxviruses. They offer cutting-edge techniques for the construction and characterization of recombinant viruses; for the study of poxvirus gene transcription and DNA replication; and for the use of the virus in immunologic and cell-cell fusion assays. Other methods cover poxvirus bioinformatics, aspects of viral pathogenesis at both a protein and an animal model level, and the study of immune responses to poxvirus-the latter critical to the important role of vaccinia virus in smallpox vaccination and its potential role as a vaccine vector directed against infectious agents and cancer. The protocols presented follow the successful Methods in Molecular Biology(TM) series format, each one offering step-by-step laboratory instructions, an introduction outlining the principle behind the technique, lists of equipment and reagents, and tips on troubleshooting and avoiding known pitfalls.Comprehensive and timely, Vaccinia Virus and Poxvirology: Methods and Protocols offers virologists and vaccinologists an unequaled source of proven techniques for understanding poxviruses and developing defenses against their use as bioterrorist agents.

Working Safely with Vaccinia Virus.- Construction and Isolation of Recombinant Vaccinia Virus Using Genetic Markers.- Construction of Recombinant Vaccinia Virus.- Transient and Inducible Expression of Vaccinia/T7 Recombinant Viruses.- Construction of Recombinant Vaccinia Viruses Using Leporipoxvirus-Catalyzed Recombination and Reactivation of Orthopoxvirus DNA.- Construction of cDNA Libraries in Vaccinia Virus.- Construction and Isolation of Recombinant MVA.- Growing Poxviruses and Determining Virus Titer.- Rapid Preparation of Vaccinia Virus DNA Template for Analysis and Cloning by PCR.- Orthopoxvirus Diagnostics.- An In Vitro Transcription System for Studying Vaccinia Virus Early Genes.- An In Vitro Transcription System for Studying Vaccinia Virus Late Genes.- Studying Vaccinia Virus RNA Processing In Vitro.- Methods for Analysis of Poxvirus DNA Replication.- Studying the Binding and Entry of the Intracellular and Extracellular Enveloped Forms of Vaccinia Virus.- Pox, Dyes, and Videotape.- Interaction Analysis of Viral Cytokine-Binding Proteins Using Surface Plasmon Resonance.- Monitoring of Human Immunological Responses to Vaccinia Virus.- Vaccinia Virus as a Tool for Immunologic Studies.- Mouse Models for Studying Orthopoxvirus Respiratory Infections.- Viral Glycoprotein-Mediated Cell Fusion Assays Using Vaccinia Virus Vectors.- Use of Dual Recombinant Vaccinia Virus Vectors to Assay Viral Glycoprotein-Mediated Fusion with Transfection-Resistant Primary Cell Targets.- Poxvirus Bioinformatics.- Preparation and Use of Molluscum Contagiosum Virus from Human Tissue Biopsy Specimens.