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

Recommendations

Info

Chapter 11 Studying Gene Expression and Function 201 Figure 11.21 Hybridized mRNA cannot be translated Protein coded by the cDNA Gene Properties of the normal protein mRNA preparation Protein Add specific cDNA Cell-free translation HART products Total translation products Compare cDNA hybridizes to the mRNA counterpart Gel electrophoresis, autoradiography Mutation, e.g. T A * Properties of the mutated protein Figure 11.20 Hybrid-arrest translation. * Mutation, e.g. ile leu A mutation may change the amino acid sequence of a protein, possibly affecting its properties. protein and its mode of activity. The best way of tackling these problems is to induce a mutation in the gene coding for the protein and then to determine what effect the change in amino acid sequence has on the properties of the translation product (Figure 11.21). Under normal circumstances mutations occur randomly and a large number may have to be screened before one that gives useful information is found. A solution to this problem is provided by in vitro mutagenesis, a technique that enables a directed mutation to be made at a specific point in a cloned gene.
Page 4:
GENE CLONING AND DNA ANALYSIS
Page 10:
This edition first published 2010,
Page 16:
Contents CONTENTS Preface to the Si
Page 20:
Contents ix 4.3 Ligation—joining
Page 24:
Contents xi 8 How to Obtain a Clone
Page 28:
Contents xiii 11.3 Identifying and
Page 32:
Contents xv 15.1.2 Herbicide resist
Page 36:
PART I The Basic Principles of Gene
Page 42:
4 Part I The Basic Principles of Ge
Page 46:
6 1.4 What is PCR? Figure 1.2 The b
Page 50:
8 Figure 1.3 Cloning allows individ
Page 54:
10 Figure 1.5 Gene isolation by PCR
Page 58:
12 Further reading FURTHER READING
Page 62:
14 Figure 2.1 Plasmids: independent
Page 66:
16 Figure 2.4 Plasmid transfer by c
Page 70:
18 Figure 2.6 Capsid components Pha
Page 74:
20 Figure 2.7 λ phage particle att
Page 78:
22 (a) The linear form of the λ DN
Page 82:
24 Part I The Basic Principles of G
Page 86:
26 Bacterial culture Table 3.1 Cent
Page 90:
28 Bacterial culture Figure 3.3 Har
Page 94:
30 Figure 3.6 Removal of protein co
Page 98:
32 Figure 3.8 Collecting DNA by eth
Page 102:
34 Figure 3.10 DNA purification by
Page 106:
36 Figure 3.12 Two conformations of
Page 110:
38 Figure 3.15 Partial unwinding of
Page 114:
40 Figure 3.18 Preparation of a pha
Page 118:
42 Figure 3.21 Collection of phage
Page 122:
44 Further reading FURTHER READING
Page 126:
Page 130:
48 Figure 4.3 The reactions catalyz
Page 134:
50 Figure 4.6 (a) Alkaline phosphat
Page 138:
52 Figure 4.8 (a) Restriction of ph
Page 142:
54 Figure 4.9 (a) Production of blu
Page 146:
56 Table 4.2 A 10 × buffer suitabl
Page 150:
58 Figure 4.13 Visualizing DNA band
Page 154:
60 Figure 4.15 Using a restriction
Page 158:
62 Figure 4.17 The influence of DNA
Page 162:
64 Figure 4.20 (a) Ligating blunt e
Page 166:
66 Figure 4.23 Adaptors and the pot
Page 170:
68 Figure 4.25 The use of adaptors:
Page 174:
70 Figure 4.28 (a) The ends of the
Page 178:
72 Chapter 5 Introduction of DNA in
Page 182:
Page 186:
76 Figure 5.4 Normal E. coli cell (
Page 190:
78 Figure 5.8 (b) Replica plating W
Page 194:
80 Figure 5.10 (a) The role of the
Page 198:
82 Figure 5.11 (a) A single-strain
Page 202:
84 Figure 5.13 Strategies for the s
Page 206:
86 Figure 5.14 Figure 5.15 (a) Prec
Page 210:
88 Chapter 6 Cloning Vectors for E.
Page 214:
90 during purification. pBR322 is 4
Page 218:
92 (a) pUC8 (b) Restriction sites i
Page 222:
94 Figure 6.5 The M13 genome, showi
Page 226:
96 not totally disrupt the lacZ′
Page 230:
98 Figure 6.10 The λ genetic map,
Page 234:
100 (a) Cloning with a λ replaceme
Page 238:
102 Figure 6.15 (a) A typical cosmi
Page 242:
104 capsid structure. Cosmid-type v
Page 246:
106 Figure 7.1 The yeast 2 µm plas
Page 250:
108 Figure 7.4 Cloning with an E. c
Page 254:
110 Figure 7.7 Chromosome structure
Page 258:
112 for instance to examine the seg
Page 262:
114 Figure 7.10 The Ti plasmid and
Page 266:
116 (a) Wound infection by recombin
Page 270:
118 Figure 7.15 Direct gene transfe
Page 274:
120 Caulimovirus vectors Although o
Page 278:
122 Figure 7.18 Cloning in Drosophi
Page 282:
124 cause the death of the mouse ce
Page 286:
126 Chapter 8 How to Obtain a Clone
Page 290:
128 Figure 8.2 The basic strategies
Page 294:
130 Figure 8.4 (a) Construction of
Page 298:
132 Figure 8.5 Preparation of a gen
Page 302:
134 Figure 8.7 One possible scheme
Page 306:
136 Figure 8.10 The structure of α
Page 310:
138 Figure 8.12 Two methods for the
Page 314:
140 Figure 8.15 A simplified scheme
Page 318:
142 Figure 8.17 Heterologous probin
Page 322:
144 skills (Figure 8.19b). The same
Page 326:
146 The problem of gene expression
Page 330:
148 Figure 9.1 Hybridization of the
Page 334:
150 Figure 9.3 Figure 9.4 5’ 3’
Page 338:
152 Figure 9.7 A typical temperatur
Page 342:
154 Figure 9.9 Calculating the T m
Page 346:
156 Figure 9.13 5’ 3’ A G A C T
Page 350:
158 Figure 9.16 Double-stranded PCR
Page 354:
160 Figure 9.18 Hybridization of a
Page 360:
PART II The Applications of Gene Cl
Page 366:
166 termination sequencing has gain
Page 370:
168 Figure 10.3 Reading the sequenc
Page 374:
170 Figure 10.5 The basis to therma
Page 378:
172 Figure 10.7 Pyrosequencing. Fig
Page 382:
174 Figure 10.9 Part II The Applica
Page 386: 176 Figure 10.11 Using oligonucleot
Page 390: 178 Figure 10.13 Chromosome walking
Page 394: 180 Figure 10.16 Part II The Applic
Page 398: 182 Figure 10.20 Fluorescent in sit
Page 402: 184 s Part II The Applications of G
Page 406: 186 Figure 11.1 Some fundamentals o
Page 410: 188 Figure 11.3 The 5′ end of an
Page 414: 190 Figure 11.6 5' 3' RNA transcrip
Page 418: 192 Figure 11.8 Possible positions
Page 422: 194 Figure 11.11 A bound protein pr
Page 426: 196 Figure 11.14 A modification int
Page 430: 198 Table 11.1 Figure 11.16 A repor
Page 434: 200 Pure mRNA Labelled translation
Page 440: Chapter 11 Studying Gene Expression
Page 444: Chapter 11 Studying Gene Expression
Page 448: Chapter 12 Studying Genomes Chapter
Page 452: Chapter 12 Studying Genomes 209 Fig
Page 460: Chapter 12 Studying Genomes 213 (a)
Page 464: Chapter 12 Studying Genomes 215 12.
Page 468: Chapter 12 Studying Genomes 217 C G
Page 480: PART III The Applications of Gene C
Page 486:
226 Figure 13.1 Two different syste
Page 490:
228 Figure 13.3 The three most impo
Page 494:
230 Figure 13.6 Strong and weak pro
Page 498:
232 Part III The Applications of Ge
Page 502:
234 Figure 13.12 Fusion protein bou
Page 506:
236 organism has a bias toward pref
Page 510:
238 Figure 13.16 Part III The Appli
Page 514:
240 Figure 13.18 Crystalline inclus
Page 518:
242 Figure 13.20 Recombinant protei
Page 522:
244 s Part III The Applications of
Page 526:
246 14.1.1 Recombinant insulin Figu
Page 530:
248 Figure 14.2 The synthesis of re
Page 534:
250 Figure 14.5 (a) The factor VIII
Page 538:
252 Figure 14.7 Part III The Applic
Page 542:
Page 546:
Page 550:
258 Figure 14.10 Part III The Appli
Page 554:
260 Figure 14.11 Differentiation of
Page 558:
Page 562:
264 Chapter 15 Gene Cloning and DNA
Page 566:
266 Table 15.1 Part III The Applica
Page 570:
268 Figure 15.3 Positional effects.
Page 574:
270 (a) Production of two toxins (c
Page 578:
272 (a) Detoxification of glyphosat
Page 582:
274 are being produced by transferr
Page 586:
276 Figure 15.10 The differences in
Page 590:
278 Figure 15.11 DNA excision by th
Page 594:
Page 598:
282 Chapter 16 Gene Cloning and DNA
Page 602:
284 Figure 16.1 Genetic fingerprint
Page 606:
286 Figure 16.3 Inheritance of STR
Page 610:
Page 614:
290 Figure 16.6 Sex identification
Page 618:
Page 622:
Page 626:
296 Figure 16.11 Origin of agricult
Page 630:
298 Glossary GLOSSARY 2 Fm circle A
Page 634:
300 Glossary Chromosome walking A t
Page 638:
302 Glossary Ethanol precipitation
Page 642:
304 Glossary In vitro mutagenesis A
Page 646:
306 Glossary Nick translation The r
Page 650:
308 Glossary Proteome The entire pr
Page 654:
310 Glossary Single nucleotide poly
Page 658:
312 INDEX Index (g = glossary, f =
Page 662:
314 cloning vectors (continued) exp
Page 666:
316 herpes simplex virus glycoprote
Page 670:
318 polyethylene glycol, see PEG po
Page 674:
320 T m , 153, 305g tobacco, 269 TO
show all

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

Create successful ePaper yourself

Delete template?

Save as template?