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Herpes Simplex Virus: Methods and Protocols provides a wide collection of protocols employed in various levels of herpes virus research, including basic protocols on growing viruses in cell culture and cloning, manipulating, and preparing viral DNA. Other chapters describe approaches to design and apply HSV-1 vectors for vaccination, cancer and gene therapy, or to study specific aspects of HSV-1 biology such as latency, intracellular transport, and protein-protein interaction. Procedures for structural analyses, microscopy, proteomics, and testing of antivirals are included as well. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
1. HSV-1 Biology and Life Cycle
Roger D. Everett
2. Herpes Simplex Virus Growth, Preparation, and Assay
Peggy Marconi and Roberto Manservigi
3. Isolation of Herpes Simplex Virus Nucleocapsid DNA
Moriah Szpara
4. Construction and Characterization of Bacterial Artificial Chromosomes (BACs) Containing Herpes Simplex Virus Full-Length Genomes
Claus-Henning Nagel, Anja Pohlmann, and Beate Sodeik
5. Engineering HSV-1 Vectors for Gene Therapy
William F. Goins, Shaohua Huang, Justus B. Cohen, and Joseph C. Glorioso
6. Herpes Simplex Virus Type 1 (HSV-1)-Derived Amplicon Vectors
Matias E. Melendez, Cornel Fraefel, and Alberto L. Epstein
7. HSV-1 Amplicon Vectors as Genetic Vaccines
Andrea S. Laimbacher and Cornel Fraefel
8. Modification of HSV-1 to an Oncolytic Virus
Hiroshi Nakashima and E. Antonio Chiocca
9. Herpes Simplex Virus Mutant Generation and Dual Detection Methods for Gaining Insight into Latent/Lytic Cycles In Vivo
Nancy M. Sawtell and Richard L. Thompson
10. Phenotypic and Genotypic Testing of HSV-1 Resistance to Antivirals
Andreas Sauerbrei and Kathrin Bohn
11. Using Homogeneous Primary Neuron Cultures to Study Fundamental Aspects of HSV-1 Latency and Reactivation
Ju Youn Kim, Lora A. Shiflett, Jessica A. Linderman, Ian Mohr, and Angus C. Wilson
12. Characterization of Extracellular HSV-1 Virions by Proteomics
Roger Lippé
13. A Precipitation-Based Assay to Analyze Interactions of Viral Particles with Cytosolic Host Factors
Kerstin Radtke, Fenja Anderson, and Beate Sodeik
14. Affinity Purification Combined with Mass Spectrometry to Identify Herpes Simplex Virus Protein-Protein Interactions
David G. Meckes, Jr.
15. Preparation of Herpes Simplex Virus Infected Primary Neurons for Transmission Electron Microscopy
Monica Miranda-Saksena, Ross Boadle, and Anthony L. Cunningham
16. Multifluorescence Live Analysis of Herpes Simplex Virus Type-1 Replication
Michael Seyffert, Anna Paula de Oliveira, Cornel Fraefel, and Rebecca Vogel
17. Expression, Purification, and Crystallization of HSV-1 Glycoproteins for Structure Determination
Samuel D. Stampfer and Ekaterina E. Heldwein
18. Cryo-EM Techniques to Resolve the Structure of HSV-1 Capsid-Associated Components
Ryan H. Rochat, Corey W. Hecksel and Wah Chiu
19. In Vivo HSV-1 DNA Transport Studies Using Murine Retinal Ganglion Cells
Jolene M. Draper, Graham S. Stephenson and Jennifer H. LaVail
20. HSV-1 Protein Expression Using Recombinant Baculoviruses
Lorry M. Grady, Ping Bai and Sandra K. Weller
21. The Murine Intravaginal HSV-2 Challenge Model for Investigation of DNA Vaccines
Joshua O. Marshak, Lichun Dong, and David M. Koelle
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