E-Book Overview
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)
E-Book Content
Methods in Molecular Genetics Volume 7 Viral Gene Techniques This Page Intentionally Left Blank Methods in Molecular Genetics Edited by Kenneth W. Adolph Department of Biochemistry University of Minnesota Medical School Minneapolis, Minnesota Volume 7 Viral Gene Techniques ACADEMIC PRESS San Diego New York Boston London Sydney Tokyo Toronto Front cover photograph: Primary human astroglioma cells infected with LacZ AAV at approximately one virus per cell. The blue nucleus within certain cells indicates infection and expression of the recombinant AAV virus. Courtesy of A. W. Skulimowski and R. J. Samulski, The Gene Therapy Center, The University of North Carolina at Chapel Hill. This book is printed on acid-flee paper. @ Copyright 9 1995 by ACADEMIC PRESS, INC. All Rights Reserved. No part of this publication may be reproduced or transmitted in any form or by any means, electronic or mechanical, including photocopy, recording, or any information storage and retrieval system, without permission in writing from the publisher. A c a d e m i c Press, Inc. A Division of Harcourt Brace & Company 525 B Street, Suite 1900, San Diego, California 92101-4495 United Kingdom Edition published by Academic Press Limited 24-28 Oval Road, London NW1 7DX International Standard Serial Number: 1067-2389 International Standard Book Number: 0-12-044309-0 PRINTED IN THE UNITED STATES OF AMERICA 95 96 97 98 99 00 EB 9 8 7 6 5 4 3 2 1 Table of Contents Contributors to Volume 7 Preface Volumes in Series ix xv xii Section I Viral Vectors 1. Adeno-Associated Virus" Integrating Vectors for Human Gene Therapy A. W. Skulimowski and R. Jude Samulski 2. Techniques for Human Adenovirus Vector Construction and Characterization Mary Hitt, Andrew J. Bett, Christina L. Addison, Ludvik Prevec, and Frank L. Graham 13 3. Construction and Isolation of Recombinant Adenoviruses with Gene Replacements David J. Spector and Lorna A. Samaniego 4. Construction and Use of Recombinant Vaccinia Virus Vectors Douglas Grosenbach and Dennis E. Hruby 31 45 5. Epstein-Barr Viral Plasmid Vectors and Their Amplifiable Derivatives Ann L. Kirchmaier, Tim A. Bloss, David Mackey, and Bill Sugden 65 Section II DNA Viruses 6. Methods to Study Epstein-Barr Virus and p53 Status in Human Cells Alison J. Sinclair and Paul J. Farrell 89 7. The SV40 Minichromosome Claudia Gruss and Rolf Knippers 101 8. Site-Specific Integration of Multigenic Shuttle Plasmids into the Herpes Simplex Virus Type 1 Genome Using a Cell-Free