Methods in Molecular Genetics Vol. 7 : Viral Gene Techniques

by Adolph, Kenneth W.

• ISBN13: 9780120443093
• ISBN10: 0120443090
• Format: Hardcover
• Copyright: 1995-09-01
• Publisher: Academic Pr

Summary

Viral Gene Techniques is a practical laboratory guide to current techniques of molecular biology and genetics. The volume is concerned with methods for the analysis of viral genes and chromosomes: DNA viruses and RNA viruses including HIV are discussed. * Methods presented for ease of use and reasdy adaptation to new systems * Detailed experimental protocols included for: * Viral vectors - construction and use of DNA virus vectors (adenovirus, adeno-associated virus, vaccinia virus, Epstein - Barr virus) * DNA viruses - virus/host interactions, viral chromosomes , transcription regulation (viruses discussed include herpes simplex, hepatitis B, SV40, JC, Epstein-Barr, adenovirus) * Human Immunodeficiency Virus / retroviruses -quantitation of HIV-1 virus stock and RNA, retrovirus reverse transcription / integration, retrovirus-mediated cell fusion, use as cell lineage markers * RNA viruses - RNA virus assembly, analysis of RNA genomes, assays for RNA-binding proteins (viruses discussed include poliovirus, influenza virus, hepatitis delta virus)

Table of Contents

Viral Vectors: Adeno-Associated Virus: Integrating Vectors for Human Gene Therapy. Techniques for Human Adenovirus Vector Construction and Characterization. Construction and Isolation of Recombinant Adenoviruses with Gene Replacements. Construction and Use of Recombinant Vaccinia Virus Vectors. Epstein-Barr Viral Plasmid Vectors and Their Amplifiable Derivatives. DNA Viruses: Methods to Study Epstein-Barr Virus and P53 Status in Human Cells. The SV40 Minichromosome. Site-Specific Integration of Multigenic Shuttle Plasmids Into the Herpes Simplex Virus Type 1 (HSV. 1). Genome Using a Cell-Free Cre-Lax Recombination System. Strategies Transcription Regulation of Neurotropic Viruses in CNS: Lesson from JC Virus. Analysis of the Nuclear DNA-Binding Activity in Cells Overexpressing Hepatitus B Viral X and 3'-Truncated S Transactivators. Investigations on Virus-Host Interations: An Abortive Systems. Human Immunodeficiency Virus/Retroviruses: Quantitation of Virus Stocks Produced from Cloned Human Immunodeficiency Virus DNA. Quantitation of HIV-1 RNA in Plasma or Serum Samples. Detection of Cell Fusion Mediated By the Envelopes of Human Retroviruses By Transactivation of a Reporter Gene. Assembly of Recombinant Retroviral Gag Precursors Into Pseudovirions in the Baculovirus-Insect Cell Expression System. Retrovirus Reverse Transcription and Integration. In Vitro Binding and Transcription Assays Using the Human T-Cell Leukemia Type I Tax Protein. PCR-Based Techniques for Utilizing Retroviruses as Cell Lineage Markers. RNA Viruses: New Methods to Study Poliovirus Assembly and Encapsidation of Genomic RNA. Molecular Studies of Hepatitis Delta Virus Rnas. Systems to Express Recombinant RNA Molecules By the Influenza a Virus Polymerase in Vivo. Analysis of Membrane -Association in Vivo and Targeting of Viral Proteins in Polarized Epithelial Cells. Cloning of Viral Dsrna Genomes By Single Primer Amplification. Using the RNA-Capture Assay to Assess the RNA-Binding Activity of Viral Proteins. Protein RNA-Binding Activity Measured By Northwestern Blot Analysis: The Interferon-Inducible, RNA-Dependent Protein Kinase PKR. Index.