Gene Cloning

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Gene Cloning Julia Lodge, Pete Lund & Steve Minchin School of Biosciences University of Birmingham Edgbaston Birmingham UK
Published by: Taylor & Francis Group In US: In UK: 270 Madison Avenue New York, N Y 10016 2 Park Square, Milton Park Abingdon, OX14 4RN This edition published in the Taylor & Francis e-Library, 2007. “To purchase your own copy of this or any of Taylor & Francis or Routledge’s collection of thousands of eBooks please go to www.eBookstore.tandf.c © 2007 by Taylor & Francis Group ISBN 0-203-96728-3 Master e-book ISBN ISBN: 0-7487-6534-4 (Print Edition) This book contains information obtained from authentic and highly regarded sources. Reprinted material is quoted with permission, and sources are indicated. A wide variety of references are listed. Reasonable efforts have been made to publish reliable data and information, but the author and the publisher cannot assume responsibility for the validity of all materials or for the consequences of their use. All rights reserved. No part of this book may be reprinted, reproduced, transmitted, or utilized in any form by any electronic, mechanical, or other means, now known or hereafter invented, including photocopying, microfilming, and recording, or in any information storage or retrieval system, without written permission from the publishers. A catalog record for this book is available from the British Library. Library of Congress Cataloging-in-Publication Data Lodge, Julia. Gene cloning : principles and applications / Julia Lodge, Pete Lund & Steve Minchin. p. ; cm. Includes bibliographical references and index. ISBN 0-7487-6534-4 (alk. paper) 1. Molecular cloning. I. Lund, Peter A. II. Minchin, Steve. III. Title. [DNLM: 1. Cloning, Molecular. 2. Gene Library. 3. Genomics --methods. QU 450 L822g 2007] QH442.2.L63 2007 660.5′5--dc22 Editor: Editorial Assistant: Production Editor: Elizabeth Owen Kirsty Lyons Georgina Lucas Taylor & Francis Group, an informa business Visit our web site at http://www.garlandscience.com
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Page 7 and 8: vi Contents 3.14 Designing PCR Prim
Page 9 and 10: viii Contents 8.3 Identifying Eukar
Page 12 and 13: 1 Introduction 1.1 The Beginning of
Page 14 and 15: Introduction 3 which also conveys a
Page 16 and 17: Introduction 5 thought, or the brin
Page 18 and 19: 2 Genome Organization Learning outc
Page 20 and 21: Genome Organization 9 and not quite
Page 22 and 23: Genome Organization 11 both of whic
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Page 26 and 27: Genome Organization 15 Box 2.2 Inte
Page 28 and 29: Genome Organization 17 Table 2.2 Hu
Page 30 and 31: Genome Organization 19 close to oth
Page 32 and 33: Genome Organization 21 individual g
Page 34 and 35: slower rate than junk DNA. (To be m
Page 36 and 37: Genome Organization 25 kb 0 2 4 6 8
Page 38 and 39: Genome Organization 27 cloning pred
Page 40 and 41: Genome Organization 29 (a) 1 2 3 4
Page 42 and 43: Genome Organization 31 interphase a
Page 44 and 45: Genome Organization 33 A2.3. Chromo
Page 46 and 47: 3 Key Tools for Gene Cloning Learni
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Page 50 and 51: Key Tools for Gene Cloning 39 Box 3
Page 52 and 53: Key Tools for Gene Cloning 41 (a) (
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Key Tools for Gene Cloning 43 then,
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Key Tools for Gene Cloning 45 Figur
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Key Tools for Gene Cloning 47 Clear
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Key Tools for Gene Cloning 49 that
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Key Tools for Gene Cloning 51 same
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3.9 More About Vectors Detecting su
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Key Tools for Gene Cloning 55 in th
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Key Tools for Gene Cloning 57 Q3.13
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Key Tools for Gene Cloning 59 alway
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Key Tools for Gene Cloning 61 Figur
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Key Tools for Gene Cloning 63 (a) C
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Key Tools for Gene Cloning 65 Box 3
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Key Tools for Gene Cloning 67 simpl
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Key Tools for Gene Cloning 69 (a) 5
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Key Tools for Gene Cloning 71 92 92
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Key Tools for Gene Cloning 73 polym
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Key Tools for Gene Cloning 75 (a) T
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Key Tools for Gene Cloning 77 DNA s
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Key Tools for Gene Cloning 79 Q3.7.
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Key Tools for Gene Cloning 81 A3.15
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Key Tools for Gene Cloning 83 A3.20
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86 Gene Cloning you extract DNA fro
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88 Gene Cloning Box 4.1 Preparation
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90 Gene Cloning which is 6.3 Mb, to
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92 Gene Cloning Also, if all the fr
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94 Gene Cloning (a) (b) Bacterial s
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96 Gene Cloning (a) Insertion vecto
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98 Gene Cloning The strictly define
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100 Gene Cloning BamHI Ap r Tet r p
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102 Gene Cloning parB Cm r parA rep
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104 Gene Cloning (a) TTTTT AAAAA TT
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106 Gene Cloning Box 4.3 Converting
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108 Gene Cloning of your gene is. A
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110 Gene Cloning A4.2. Extract chro
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112 Gene Cloning A4.8. See Section
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114 Gene Cloning Q4.15. What are th
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116 Gene Cloning dystrophin gene is
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118 Gene Cloning it is still a form
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120 Gene Cloning One obvious proble
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122 Gene Cloning The main difficult
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124 Gene Cloning (a) Labeled single
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126 Gene Cloning N terminus Phe Val
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128 Gene Cloning Box 5.3 Types of D
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130 Gene Cloning Box 5.4 Methods fo
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132 Gene Cloning the DNA probe. Bot
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134 Gene Cloning a b c d e f g h 1
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136 Gene Cloning S. cerevisiae, and
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138 Gene Cloning Q5.7. Would you ex
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140 Gene Cloning use your genomic c
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142 Gene Cloning diseases, where th
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144 Gene Cloning (a) Replicative Ta
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146 Gene Cloning where the transpos
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148 Gene Cloning DNA fragment in th
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150 Gene Cloning Bacteria transform
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152 Gene Cloning Wild-type gene Mut
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154 Gene Cloning molecule using PCR
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156 Gene Cloning digesting chromoso
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158 Gene Cloning + + Ac Avr Avr cf-
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160 Gene Cloning which are very clo
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162 Gene Cloning Box 6.3 Restrictio
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164 Gene Cloning Initial region clo
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166 Gene Cloning Box 6.4 Nonsense S
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168 Gene Cloning 6.10 Cloning of th
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170 Gene Cloning are two possible o
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172 Gene Cloning Isolation of the t
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174 Gene Cloning Two approaches wer
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176 Gene Cloning synthesize DNA de
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178 Gene Cloning Box 7.1 Denaturing
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180 Gene Cloning G A T C Figure 7.4
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182 Gene Cloning although other con
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184 Gene Cloning A large DNA fragme
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186 Gene Cloning chain reaction, fl
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188 Gene Cloning rounds of amplific
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190 Gene Cloning Contig 2 Clone 12
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192 Gene Cloning λ Clone Scaffold
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194 Gene Cloning Genomic DNA (a) Cl
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196 Gene Cloning therefore sequence
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198 Gene Cloning Tag sequence CGTGT
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200 Gene Cloning Genomic DNA (a) Ad
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202 Gene Cloning bases. The light s
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204 Gene Cloning A7.3. (a) The dide
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8 Bioinformatics Learning outcomes:
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Bioinformatics 209 Table 8.1 The ge
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Bioinformatics 211 5' accgcgcatggtg
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Bioinformatics 213 Correlation Scor
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Bioinformatics 215 AGG T R A G T C
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Bioinformatics 217 conjunction with
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Bioinformatics 219 (c) V S V K L Q
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Bioinformatics 221 Tiny P Small Ali
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Bioinformatics 223 Matrix: EBLOSUM6
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Bioinformatics 225 Box 8.2 DNA and
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Bioinformatics 227 (a) DB:ID Source
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Bioinformatics 229 (a) Sequences pr
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Bioinformatics 231 list is also of
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Bioinformatics 233 this relates to
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Bioinformatics 235 common functiona
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Bioinformatics 237 These two regula
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Bioinformatics 239 three-dimensiona
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Bioinformatics 241 Neisseria mening
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Bioinformatics 243 A8.1. The triple
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Bioinformatics 245 Q8.11. Which gro
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Bioinformatics 247 EMBL Nucleotide
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250 Gene Cloning What sorts of stud
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252 Gene Cloning 9.2 Requirements f
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254 Gene Cloning (a) The lac promot
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256 Gene Cloning lac: CCAGGCTTTACAC
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258 Gene Cloning phage (such as T7)
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260 Gene Cloning 9.4 Some Problems
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262 Gene Cloning Figure 9.6 Inclusi
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264 Gene Cloning Box 9.1 Translatio
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266 Gene Cloning Box 9.2 Signal Seq
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268 Gene Cloning Ribosome Cytosol P
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270 Gene Cloning Other yeast specie
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272 Gene Cloning (turned on by an i
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274 Gene Cloning 9.6 A Final Word A
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276 Gene Cloning study, as they wil
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10 Gene Cloning in the Functional A
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Gene Cloning in the Functional Anal
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316 Gene Cloning genomic sequences
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318 Gene Cloning the first exon of
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320 Gene Cloning Box 11.1 End Label
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322 Gene Cloning mRNA 5' 3' 3' 5' S
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324 Gene Cloning mRNA 5' 3' 3' 5' G
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326 Gene Cloning cloning step. RACE
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328 Gene Cloning Prepare RNA from c
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330 Gene Cloning Gene A condition 1
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332 Gene Cloning (a) (b) (c) Immobi
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334 Gene Cloning tat and one in whi
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336 Gene Cloning Box 11.3 Examples
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338 Gene Cloning (a) MCS for promot
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340 Gene Cloning important regulato
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342 Gene Cloning (a) F DP DPA DNA-t
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344 Gene Cloning DNA alone DNA-prot
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346 Gene Cloning [FNR] GA - + -90 -
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348 Gene Cloning (a) 1 2 3 4 5 Prom
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350 Gene Cloning bind the transcrip
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352 Gene Cloning Target sequence Ye
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354 Gene Cloning have been develope
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356 Gene Cloning detected by autora
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358 Gene Cloning vide a complete li
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360 Gene Cloning MS 503 750 1400 16
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362 Gene Cloning Q11.3. Why is it e
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364 Gene Cloning (transcriptomics o
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366 Gene Cloning Figure 12.1 Transg
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368 Gene Cloning Herbicide Non-tran
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370 Gene Cloning that has been used
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372 Gene Cloning tumors with high f
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374 Gene Cloning Transgene encoding
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376 Gene Cloning placed on the mark
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378 Gene Cloning Gene A: a gene whi
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380 Gene Cloning Isolate the gene o
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382 Gene Cloning Box 12.1 Recombina
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384 Gene Cloning damage to the DNA
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386 Gene Cloning Box 12.2 The Produ
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388 Gene Cloning as to whether the
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390 Gene Cloning transformed. The i
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392 Gene Cloning 1. Gene of interes
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394 Gene Cloning Showing that the D
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396 Gene Cloning complete plants by
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398 Gene Cloning 12.5 Knockout Mice
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400 Gene Cloning introduced back in
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402 Gene Cloning Chromosome Constru
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404 Gene Cloning cells, and one has
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406 Gene Cloning mapped by inverse
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408 Gene Cloning traditional method
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410 Gene Cloning Further Reading Th
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412 Gene Cloning (a) Maternal chrom
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414 Gene Cloning chromosomes togeth
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416 Gene Cloning Box 13.1 DNA Profi
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418 Gene Cloning Although forensic
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420 Gene Cloning Dye terminators (a
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422 Gene Cloning With the advent of
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424 Gene Cloning Box 13.2 Genetic A
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426 Gene Cloning Box 13.3 Methods o
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428 Gene Cloning (a) PCR primer seq
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430 Gene Cloning (a) Target specifi
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432 Gene Cloning (a) R Q 5' 3' (b)
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434 Gene Cloning liquid chromatogra
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436 Gene Cloning such as pre-implan
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438 Gene Cloning attack there is a
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440 Gene Cloning and also because l
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442 Gene Cloning John Mary Ann Pete
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444 Gene Cloning monitoring the pro
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446 Gene Cloning Codon Three nucleo
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448 Gene Cloning Homologous recombi
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450 Gene Cloning Phenotype The obse
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452 Gene Cloning Vector A self-repl
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454 Gene Cloning Capillary electrop
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456 Gene Cloning EST see expressed
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458 Gene Cloning Metallothionein, 3
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460 Gene Cloning Protein addition o
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462 Gene Cloning Transcription, ide
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Gene Cloning

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