Coronavirus Replication and Reverse Genetics

Coronaviruses are the RNA viruses with the largest genome known to date (27 to 32 kb). Members of this virus family affect most domestic animal species, causing important socio-economical losses, and also infect humans. Human coronaviruses were known to c

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Editors R.W. Compans, Atlanta/Georgia M.D. Cooper, Birmingham/Alabama T. Honjo, Kyoto · H. Koprowski, Philadelphia/Pennsylvania F. Melchers, Basel · M.B.A. Oldstone, La Jolla/California S. Olsnes, Oslo · M. Potter, Bethesda/Maryland P.K. Vogt, La Jolla/California · H. Wagner, Munich

L. Enjuanes (Ed.)

Coronavirus Replication and Reverse Genetics With 49 Figures

Professor Dr. Luis Enjuanes Centro Nacional de Biotecnologa Department of Molecular and Cell Biology Campus Universidad Autnoma, Cantoblanco 38049 Madrid Spain e-mail: [email protected] Cover illustration by Luis Enjuanes Simplified structure of a coronavirus prototype (transmissible gastroenteritis virus, TGEV) showing the envelope with several structural proteins: spike (S), two topologies of the membrane (M or M') proteins, envelope (E) protein, and nucleoprotein (N). A nucleocapsid can be observed inside the envelope virus, containing the genomic RNA, the N protein, and the carboxiterminus of the M protein.

Library of Congress Catalog Card Number 72-152360 ISSN 0070-217X ISBN 3-540-21494-1 Springer Berlin Heidelberg New York This work is subject to copyright. All rights are reserved, whether the whole or part of the material is concerned, specifically the rights of translation, reprinting, reuse of illustrations, recitation, broadcasting, reproduction on microfilms or in any other way, and storage in data banks. Duplication of this publication or parts thereof is permitted only under the provisions of the German Copyright Law of September 9, 1965, in its current version, and permission for use must always be obtained from Springer-Verlag. Violations are liable for prosecution under the German Copyright Law. Springer is a part of Springer Science+Business Media springeronline.com  Springer-Verlag Berlin Heidelberg 2005 Printed in Germany The use of general descriptive names, registered names, trademarks, etc. in this publication does not imply, even in the absence of a specific statement, that such names are exempt from the relevant protective laws and regulations and therefore free for general use. Product liability: The publishers cannot quarantee that accuracy of any information about dosage and application contained in this book. In every individual case the user must check such information by consulting the relevant literature. Editor: Dr. Rolf Lange, Heidelberg Desk editor: Anne Clauss, Heidelberg Production editor: Andreas Gsling, Heidelberg Cover design: design & production GmbH, Heidelberg Typesetting: Strtz AG, Wrzburg Printed on acid-free paper 27/3150/ag – 5 4 3 2 1 0

Preface

The Coronaviridae family is included in the Nidovirales order together with the Arteriviridae and Roniviridae. Possibly the first recorded coronavirus-related disease was feline infectious peritonitis in 1912. However, until the late 1960s the coronaviruses were not recognized as pathogens responsible for human diseases (common cold), and it was in 2003 when human coronaviruses (HCoVs) received worldwide attention with the emergence of the severe and acu