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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 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.A comprehensive collection of cutting-edge methods to study and work with the vaccinia virus and other poxvirus gene transcriptions. These readily reproducible techniques can be used for the construction and characterization of recombinant viruses; for the study of poxvirus gene transcription and DNA replication; for the investigation of the binding, entry, and movement of the virus in host cells; 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 poxviruses-the latter a critical ability given the important role of vaccinia virus in smallpox vaccination and their potential role as vaccine vectors directed against infectious agents and cancer.