GENE CLONING & DNA ANALYSIS Vem altaelelereiteya) alee Oo= Ales© 2001 by Blackwell Scienee Lid © 2006 T.A. Brown Blackwell Publishing Editorial offices: Blackwell Publishing Ltd, 9600 Garsington Road, Oxford OX4 2DQ, UK “Tel: +4 (0)1865 76868, Blackwell Publishing Inc., 950 Main Street, Malden, MA 02148-5020, USA “Fel: +1 781 388 8250 Blackwell Publishing Asia Pry Ltd, $50 Swanston Street, Carlton, Victoria 3053, Australia “Tel: +61 (0)3 8359 1011 “The right of the Author to be identified as the Author of this Work has been asserted in accordance with the Copyright, Designs and Patents Act 1988. All rights reserved. No part ofthis publication may be reproduced. stored ina retrieval system, fr transmitted, in any form ot by any means, electronic, mechanical, photocopying, recording oF btherwise, except as permitted by the UK Copyright, Designs and Patents Act 1988, prior permission of the publisher. First, second and third editions published by Chapman & Hall 1986, 1990, 1995 Reprinted 1998 by Stanley Thomes (Publishers) Ltd Fourth edition published by Blackwell Science Lt 2001 Filth edition published 2006 SBN-13:978-14051-1121-8 ISBN-10: 1-401-1121-6 Library of Congress Cataloging: Brown, T.A. (Terence A.) Gene cloning and DNA analysis: an introduction /"EA. Brown.~ Sth ed Includes bibliographical references and index. ISBN- 13: 978-1-4051-1121-8 (pbk. alk. paper) ISBN- 10: 1-4051-1121-6 (pbk. : alk. paper) 1. Molecular cloning. 2. Nucleotide sequence. 3. DNA — Analysis IDNLM: 1. Cloning, Molecular. 2. DNA, Recombinant ~ analysis 3. Sequence Analysis DNA. OH 42.2 BS8tg 2005] 1. Tite. OHHE2.B%. 2108 572.8" — de22 20005006606 ‘A catalogue record for this title wailable from the British Library Set in 10/13 PUTimes by SNP Best-set Typesetter Lid, Hong Kong. Printed and bound in India bby Replika Press Pvt, Lid, Kundli ‘The publisher's policy is to use permanent paper from mill that operate a sustainable forestry policy, and which has been manufactured {rom pulp processed using acid-free and elementary Chlorine-free practices. Furthermore, the publisher ensures that the text paper and cover board ‘used have met acceptable enviroomental accreditation standards For further information on Blackwell Publishing, vist our website: www blackwellpublishing comContents Preface to the Fifth Edition, xv Preface to the Fourth Edition, xvi Preface to the Third Edition, xvii Preface to the Second Edition, xviii Preface to the First Edition, x Part 1: The Basic Principles of Gene Cloning and DNA Analysis, 1 1_Why Gene Cloning and DNA Analysis are Important, 3 1.1 __ The early development of genetics. 3 1.2 The advent of gene cloning and the polymerase chain reaction, 4 1.3. What is gene cloning? 5 14 What is PCR? 6 1.5 Why gene cloning and PCR are so important. 8 15.1 Gene isolation by cloning, 8 15.2 Gene isolation by PCR, 9 1.6 How to find your way through this book, 12 2 Vectors for Gene Cloning: Plasmids and Bacteriophages, 14 21 Plasmids, 14 21.1 Basic features of plasmids, 14 21.2 Size and copy number, 16 2.1.3. Conjugation and compatibility, 17 4 _ Plasmid classification, 18 2.1.5 Plasmids in organisms other tha 2.2 Bacteriophages, 19 2.2.1 Basic features of bacteriophages, 19 2.2.2 Lysogenic phages, 20 bacteria, 19