Adenovirus Methods and Protocols: Ad Proteins and RNA, Lifecycle and Host Interactions, and Phyologenetics

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William S. M. Wold, Ann E. Tollefson
Springer Science & Business Media, 2006 - Science - 698 pages
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Adenovirus Methods and Protocols, Second Edition, now in two volumes, is an essential resource for adenovirus (Ad) researchers beginning in the field, and an inspirational starting point for researchers looking to branch into new areas of Ad study. In addition to updating and expanding important chapters from the first edition, the authors have added new chapters that address innovative, exciting areas of emphasis in Ad research, including Ad vector construction and use, real-time PCR, use of new animal models, and methods for quantification of Ad virus or virus expression/interactions. Each of the protocols presented in these volumes is written by trendsetting researchers in their respective areas of expertise. Volume 1 addresses several important techniques for construction of adenoviruses for use as vectors and for basic research. Highlighted topics include deletion mutants, capsid modifications, insertions, and gene replacements in human, murine, bovine, and ovine adenoviruses. In Volume 2, the authors focus on methods that elucidate and quantitate the interactions of Ad with the host. Each of the protocols in these volumes provides a general introduction, followed by tried-and-true step-by-step methods. Both novice and experienced researchers will reap tremendous benefit from these groundbreaking volumes in Ad research.
 

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Contents

1 Analysis of the Efficiency of Adenovirus Transcription
1
2 The Use of In Vitro Transcription to Probe Regulatory Functions of Viral Protein Domains
15
3 Preparation of Soluble Extracts From AdenovirusInfected Cells for Studies of RNA Splicing
33
4 In Vitro Methods to Study RNA Interference During an Adenovirus Infection
47
5 Simultaneous Detection of Adenovirus RNA and Cellular Proteins by Fluorescent Labeling In Situ
63
6 Study of Nucleolar Localization of Adenovirus Core Proteins
73
7 Analysis of Adenovirus Infections in Synchronized Cells
83
8 CoImmunoprecipitation of Protein Complexes
103
14 Human Adenovirus Type 12
197
15 Measurement of NaturalKiller Cell Lytic Activity of AdenovirusInfected or AdenovirusTransformed Cells
213
16 A Flow Cytometric Assay for Analysis of NaturalKiller CellMediated Cytolysis of AdenovirusTransformed Cells
221
17 LargeScale Purification and Crystallization of Adenovirus Hexon
231
18 Synthesis and Assay of Recombinant Adenovirus Protease
251
19 Assay for the Adenovirus Proteinase
257
20 Cofactors of the Adenovirus Proteinase
269
21 Characterization of the Adenovirus Fiber Protein
281

9 Chromatin Immunoprecipitation to Study the Binding of Proteins to the Adenovirus Genome In Vivo
113
10 Assaying ProteinDNA Interactions In Vivo and In Vitro Using Chromatin Immunoprecipitation and Electrophoretic Mobility Shift Assays
123
11 Identifying Functional AdenovirusHost Interactions Using Tandem Mass Spectrometry
141
12 The Use of Cell Microinjection for the In Vivo Analysis of Viral Transcriptional Regulatory Protein Domains
157
13 Determination of the Transforming Activities of Adenovirus Oncogenes
187
22 Phylogenetic Analysis of Adenovirus Sequences
299
23 Assessment of Genetic Variability Among Subspecies B1 Human Adenoviruses for Molecular Epidemiology Studies
335
Index
357
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