Adenovirus Methods and Protocols

A collection of readily-reproducible methods for conducting research with adenoviruses.

A cutting-edge collection of readily reproducible methods for conducting research with adenoviruses, the premier and most widely used model in cell and molecular biology. The methods range from how to grow and titer adenoviruses and how to construct specific alterations in the adenovirus genome, to how to measure apoptosis induced by cells of the immune system, cytokines, and intrinsic apoptosis effectors. In addition, there are methods to study transcription and splicing with in vitro systems and for the adenovirus-mediated transformation of cells to a malignant state. Each method is written by a prominent investigator well-versed in the technique and includes a brief background discussion and tried, as well as true step-by-step instructions.

Preparation and Titration of CsCL-banded Adenovirus Stocks A. E. Tollefson T. W. Hermiston and W. S. M. Wold. Construction of Mutations in the Adenovirus Early Region 3 (E3) Transcription Unit T. E. Hermiston A. E. Tollefson and W. S. M. Wold. Isolation Growth and Purification of Defective Adenovirus Deletion Mutants G. Ketner and J. Boyer. Manipulation of Early Region 4 J. Boyer and G. Ketner. Adenovirus DNA Packaging-Construction and Analysis of Viral Mutants S. I. Schmid and P. Hearing. Methods for Creating and Analyzing Adenovirus Vectors Which Express Proteins that Act on the Viral Genome C. S. H. Young A. L. Nicolas H. Lu and P. L. Munz. Construction of Mouse Adenovirus Type 1 Mutants T. E. Sparer and L. R. Gooding. Measurement of Macrophage-Mediated Cytolysis of Adenovirus-Infected Cells P. J. Duerkson- Hughes and L. R. Gooding. Measurement of TNF (Tumor Necrosis Factor) Lysis of Adenovirus- Infected Cells J. Shisler and L. R. Gooding. Role of NK Cells in Immunity to Adenoviruses J. M. Routes. Determination of the Transforming Activities of Adenovirus Oncogenes T. Subramanian and G. Chinnadurai Cell Survival and Apoptosis Assays for E1B 19K and Bcl-2 Family Members T. Subramanian and G. Chinnadurai In Vitro Transcription: Probing the Molecular Functions of Adenovirus Regulatory Proteins P. M. Lowenstein C. -Z Song and M. Green. Cell Microinjection: In Vivo Analysis of the Functional Domains of Viral Regulatory Proteins M. Green A Thornburn and P. M. Lowenstein. Immunoprecipitation of E1A-Containing Protein Complexes E. Moran and P. Yaciuk. Preparation of Splicing Competent Nuclear Extracts from Adenovirus Infected Cells O. M?hlemann and G. Akusjarvi. Adenovirus Entry Into Cells A Quantitative Fluorescence Microscopy Approach U. F. Greber M. Nakano and M. Suomalainen. Simultaneous Detection of RNA and Proteins in Adenovirus Infected Cells by Fluorescence In Situ Hybridization and Immunostaining E. Bridge. Characterization of the Adenovirus Fiber Protein J. A. Engler B. Mulach and J. S. Hong. Large Scale Purification and Crystallization of Adenovirus Hexon J. J. Rux D. Pascolini and R. M. Nurnett. Adenovirus Protease J. M. Weber. Methods for Growth and Purification of Enteric Adenovirus Type 40 V. Mautner. Transfection Complexes Generated with Adenovirus and Polyethyleneimine- Condensed DNA M. Cotten M. Saltik and A. Baker. Phylogenetic Analysis of Adenovirus Sequences: Proof of the Necessity of Establishing a Third Genus in the Adenoviridae Family B. Harrach and M. Benko. Index.