Gene Cloning and DNA Analysis: An Introduction, Sixth Edition ...

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316 herpes simplex virus glycoprotein, 254, 255t Hershey, A., 4 heterologous probing, 141–142, 303g heteroplasmy 288, 303g HindIII, 53t HinfI, 52, 53t HIV, 255t Homo erectus, 291–292 homology, 210, 303g homology search, 210–211, 303g homopolymer tailing, 67–69, 303g horseradish peroxidase, 137, 303g host-controlled restriction, 51, 303g HRT, 199–200, 303g human gene length, 208 human genome Human Genome Project, 173, 177 library, 103t, 112 size, 173, 173t human growth hormones, 247–249 human mitochondrial genome, 173 Huntington’s chorea, 255t, 255–256 hybrid protein, 232–234, 247 hybrid-arrest translation, 199–200, 303g hybridization analysis abundancy probing, 137–138 colony hybridization probing, 133–137 definition, 133 fluorescence in situ hybridization, 182 heterologous probing, 141–142 hybridization, 133 melting temperature, 153 mismatched hybrids, 152 northern hybridization, 144, 186–187, 275 nuclease analysis, 189–190 oligonucleotide probes, 138–140 plaque hybridization probing, 133–137 Southern hybridization, 142–144 uses, 137–144 hybridization probe, 133, 303g hybrid-release translation, 199–200, 303g hygromycin phosphotransferase, 198t hypervariable dispersed repetitive sequence, 283, 303g ICAT, 219 immune disorders, 251t immunological screening, 145–146, 303g in vitro mutagenesis, 98, 200–206, 304g in vitro packaging, 81, 98, 101, 304g in vitro transcription, 93–94 inclusion body, 236, 240, 304g incompatibility group, 16, 304g indel, 290, 304g induction bacteriophage, 40, 98, 304g gene, 229, 304g influenza virus, 254, 255t insect cells, 240–241 insecticides, 265–270 insertion vectors, 98–100, 304g insertional inactivation, 77–80, 83–84, 99–100, 304g insulin, 246–247, 251t intercalation, 37 interferons, 251, 251t interleukins, 242, 251, 251t interspersed repeat element PCR, 179, 304g intron, 185, 187, 190, 235–236, 250 invertase, 226t ion exchange chromatography, 30, 31, 33, 304g IPTG, 80, 231, 232 IRE-PCR, 179, 304g isoelectric focussing, 218 isoelectric point, 218 isopropyl–thiogalactoside, 80, 231, 232 isotope coded affinity tag, 219 kanamycin, 116–117, 118, 119, 269, 275, 277 kanamycin-resistance gene, safety to humans, 277 kidney transplants, 251t kinship analysis, 286–289, 304g Klebsiella ozaenae, 273t Klenow fragment, 48–49, 69, 169, 304g Kluveromyces lactis, 112, 238 knockout mouse, 124, 213, 214, 258, 304g labelling, see <strong>DNA</strong> labelling lac promoter, 231, 247–248 Lac selection, 79–80, 83, 92–93, 97, 100, 304g Lactose, 79 lacZ, 198t lacZ′, 79–80, 83, 92–93, 94–96, 97, 100, 231 Index eEMBL4, 100 egt10, 99 eP L promoter, 231–232 eZAPII, 100 late onset disease, 255–256 LB medium, 26t, 27 LEU2, 106, 109 Leuconostoc spp., 226t leukaemia, 251t ligase function, 46, 47–48, 304g mode of action, 63 ligation attaching sticky ends to a blunt–ended molecule, 64–69 products of ligation, 72–74 sticky vs blunt ends, 64 light microscopy, 182 linkage analysis, 256–257, 304g linkers, 64–65, 304g liposome, 85, 305g lithium acetate, 85 lithium chloride, 85 luciferase, 198t lung surfactant protein, 251t Luria–Bertani medium, 26t, 27 lux, 198t lysogen, 19, 232, 305g lysogenic phage, 19–24, 305g bacteriophage e, 19–21 bacteriophage M13, 22–23 lysozyme, 28–29, 32, 35, 305g lytic infection cycle, 18–19, 305g M9 medium, 26t, 26–27 M13mp2, 94–95 M13mp7, 95 M13mp8, 96 MacLeod, C., 4 maize, 266–268 major surface antigen, 252–253, 255t malaria parasite, 63 MALDI-TOF, 218–220, 305g malE, 233 mammoth genome, 173 mapping, 180–183 mapping reagent, 182, 305g marker rescue, 129–131 mass spectrometry, 217 massively parallel sequencing, 171–173, 305g matrix-assisted laser desorption ionization time-of-flight, 218–220, 305g McCarty, M., 4 measles virus, 253 mega–YAC, 112 melting temperature, 153, 305g
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GENE CLONING AND DNA ANALYSIS
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This edition first published 2010,
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Contents CONTENTS Preface to the Si
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Contents ix 4.3 Ligation—joining
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Contents xi 8 How to Obtain a Clone
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Contents xiii 11.3 Identifying and
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Contents xv 15.1.2 Herbicide resist
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PART I The Basic Principles of Gene
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4 Part I The Basic Principles of Ge
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6 1.4 What is PCR? Figure 1.2 The b
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8 Figure 1.3 Cloning allows individ
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10 Figure 1.5 Gene isolation by PCR
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12 Further reading FURTHER READING
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14 Figure 2.1 Plasmids: independent
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16 Figure 2.4 Plasmid transfer by c
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18 Figure 2.6 Capsid components Pha
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20 Figure 2.7 λ phage particle att
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22 (a) The linear form of the λ DN
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24 Part I The Basic Principles of G
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26 Bacterial culture Table 3.1 Cent
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28 Bacterial culture Figure 3.3 Har
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30 Figure 3.6 Removal of protein co
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32 Figure 3.8 Collecting DNA by eth
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34 Figure 3.10 DNA purification by
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36 Figure 3.12 Two conformations of
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38 Figure 3.15 Partial unwinding of
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40 Figure 3.18 Preparation of a pha
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42 Figure 3.21 Collection of phage
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44 Further reading FURTHER READING
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48 Figure 4.3 The reactions catalyz
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50 Figure 4.6 (a) Alkaline phosphat
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52 Figure 4.8 (a) Restriction of ph
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54 Figure 4.9 (a) Production of blu
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56 Table 4.2 A 10 × buffer suitabl
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58 Figure 4.13 Visualizing DNA band
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60 Figure 4.15 Using a restriction
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62 Figure 4.17 The influence of DNA
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64 Figure 4.20 (a) Ligating blunt e
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66 Figure 4.23 Adaptors and the pot
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68 Figure 4.25 The use of adaptors:
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70 Figure 4.28 (a) The ends of the
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72 Chapter 5 Introduction of DNA in
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76 Figure 5.4 Normal E. coli cell (
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78 Figure 5.8 (b) Replica plating W
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80 Figure 5.10 (a) The role of the
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82 Figure 5.11 (a) A single-strain
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84 Figure 5.13 Strategies for the s
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86 Figure 5.14 Figure 5.15 (a) Prec
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88 Chapter 6 Cloning Vectors for E.
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90 during purification. pBR322 is 4
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92 (a) pUC8 (b) Restriction sites i
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94 Figure 6.5 The M13 genome, showi
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96 not totally disrupt the lacZ′
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98 Figure 6.10 The λ genetic map,
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100 (a) Cloning with a λ replaceme
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102 Figure 6.15 (a) A typical cosmi
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104 capsid structure. Cosmid-type v
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106 Figure 7.1 The yeast 2 µm plas
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108 Figure 7.4 Cloning with an E. c
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110 Figure 7.7 Chromosome structure
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112 for instance to examine the seg
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114 Figure 7.10 The Ti plasmid and
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116 (a) Wound infection by recombin
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118 Figure 7.15 Direct gene transfe
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120 Caulimovirus vectors Although o
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122 Figure 7.18 Cloning in Drosophi
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124 cause the death of the mouse ce
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126 Chapter 8 How to Obtain a Clone
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128 Figure 8.2 The basic strategies
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130 Figure 8.4 (a) Construction of
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132 Figure 8.5 Preparation of a gen
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134 Figure 8.7 One possible scheme
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136 Figure 8.10 The structure of α
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138 Figure 8.12 Two methods for the
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140 Figure 8.15 A simplified scheme
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142 Figure 8.17 Heterologous probin
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144 skills (Figure 8.19b). The same
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146 The problem of gene expression
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148 Figure 9.1 Hybridization of the
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150 Figure 9.3 Figure 9.4 5’ 3’
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152 Figure 9.7 A typical temperatur
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154 Figure 9.9 Calculating the T m
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156 Figure 9.13 5’ 3’ A G A C T
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158 Figure 9.16 Double-stranded PCR
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160 Figure 9.18 Hybridization of a
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PART II The Applications of Gene Cl
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166 termination sequencing has gain
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168 Figure 10.3 Reading the sequenc
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170 Figure 10.5 The basis to therma
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172 Figure 10.7 Pyrosequencing. Fig
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174 Figure 10.9 Part II The Applica
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176 Figure 10.11 Using oligonucleot
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178 Figure 10.13 Chromosome walking
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180 Figure 10.16 Part II The Applic
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182 Figure 10.20 Fluorescent in sit
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184 s Part II The Applications of G
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186 Figure 11.1 Some fundamentals o
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188 Figure 11.3 The 5′ end of an
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190 Figure 11.6 5' 3' RNA transcrip
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192 Figure 11.8 Possible positions
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194 Figure 11.11 A bound protein pr
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196 Figure 11.14 A modification int
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198 Table 11.1 Figure 11.16 A repor
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200 Pure mRNA Labelled translation
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202 Figure 11.22 (a) Restriction fr
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204 Part II The Applications of Gen
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208 as molecular biology in silico,
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210 Figure 12.4 Part II The Applica
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212 Figure 12.7 Using comparisons b
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214 Figure 12.10 The use of a yeast
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216 Figure 12.11 Serial analysis of
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218 Load the protein sample Figure
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220 Figure 12.16 Analyzing two prot
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Chapter 13 Production of Protein fr
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Chapter 14 Gene Cloning and DNA Ana
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Page 616: Chapter 16 Gene Cloning and DNA Ana
Page 632: Glossary 299 Bacteriophage or phage
Page 636: Glossary 301 CpG island A GC-rich D
Page 640: Glossary 303 Genetic map A genome m
Page 644: Glossary 305 Liposome A lipid vesic
Page 648: Glossary 307 Pharming Genetic modif
Page 652: Glossary 309 Restriction analysis D
Page 656: Glossary 311 Transgenic Referring t
Page 660: Index 313 linear and circular forms
Page 664: Index 315 bacteriophages, 19-23 clo
Page 670: 318 polyethylene glycol, see PEG po
Page 674: 320 T m , 153, 305g tobacco, 269 TO
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