From Genes to Genomes: Concepts and Applications of DNA Technology, 2nd Edition

by Jeremy W. Dale (Univ. of Surrey, UK); Malcolm von Schantz (University of Surrey, UK)

• ISBN13: 9780470017340
• ISBN10: 0470017341
• Edition: 2nd
• Format: Paperback
• Copyright: 2007-12-01
• Publisher: Wiley-Interscience

Summary

Rapid advances in a collection of techniques referred to as gene technology, genetic engineering, recombinant DNA technology and gene cloning have pushed molecular biology to the forefront of the biological sciences. This new edition of a concise, well-written textbook introduces key techniques and concepts involved in cloning genes and in studying their expression and variation. The book opens with a brief review of the basic concepts of molecular biology, before moving on to describe the key molecular methods and how they fit together. This ranges from the cloning and study of individual genes to the sequencing of whole genomes, and the analysis of genome-wide information. Finally, the book moves on to consider some of the applications of these techniques, in biotechnology, medicine and agriculture, as well as in research that is causing the current explosion of knowledge across the biological sciences. From Genes to Genomes: Concepts and Applications of DNA Technology, Second Edition includes full two-colour design throughout and an accompanying website. Specific changes for the new edition include: Strengthening of gene to genome theme Updating and reinforcing of material on proteomics, gene therapy and stem cells More eukaryotic/mammalian examples and less focus on bacteria This textbook is must-have for all undergraduates studying intermediate molecular genetics within the biological and biomedical sciences. It is also of interest for researchers and all those needing to update their knowledge of this rapidly moving field.

Author Biography

Professor Jeremy Dale and Dr Malcolm von Schantz. School of Biomedical and Molecular Sciences, University of Surrey, Guildford, Surrey GU2 SXH, UK.

Table of Contents

Preface	p. ix
Introduction	p. 1
Basic molecular biology	p. 7
Nucleic acid structure	p. 7
What is a gene?	p. 15
Information flow: gene expression	p. 16
Gene structure and organization	p. 21
How to clone a gene	p. 25
What is cloning?	p. 25
Overview of the procedures	p. 26
Gene libraries	p. 29
Hybridization	p. 30
Polymerase chain reaction	p. 32
Extraction and purification of nucleic acids	p. 33
Detection and quantitation of nucleic acids	p. 36
Gel electrophoresis	p. 37
Cutting and joining DNA	p. 41
Restriction endonucleases	p. 41
Ligation	p. 47
Modification of restriction fragment ends	p. 53
Other ways of joining DNA molecules	p. 57
Vectors	p. 61
Plasmid vectors	p. 61
Vectors based on the lambda bacteriophage	p. 69
Cosmids	p. 78
M13 vectors	p. 79
Expression vectors	p. 81
Vectors for cloning and expression in eukaryotic cells	p. 84
Supervectors: YACs and BACs	p. 92
Summary	p. 94
Genomic and cDNA libraries	p. 95
Genomic libraries	p. 96
Growing and storing libraries	p. 105
cDNA libraries	p. 106
Random, arrayed and ordered libraries	p. 113
Finding the right clone	p. 117
Screening libraries with gene probes	p. 117
Screening expression libraries with antibodies	p. 128
Subcloning	p. 130
Characterization of plasmid clones	p. 131
Polymerase chain reaction	p. 135
The PCR reaction	p. 136
PCR in practice	p. 140
Cloning PCR products	p. 144
Long-range PCR	p. 146
Reverse-transcription PCR	p. 146
Rapid amplification of cDNA ends	p. 147
Quantitative PCR	p. 149
Applications of PCR	p. 154
Characterization of a cloned gene	p. 159
DNA sequencing	p. 159
Databank entries and annotation	p. 167
Sequence analysis	p. 173
Sequence comparisons	p. 178
Protein structure	p. 190
Confirming gene function	p. 196
Analysis of gene expression	p. 201
Analysing transcription	p. 201
Methods for studying the promoter	p. 209
Regulatory elements and DNA-binding proteins	p. 213
Translational analysis	p. 217
Products from native and manipulated cloned genes	p. 221
Factors affecting expression of cloned genes	p. 222
Expression of cloned genes in bacteria	p. 227
Expression in eukaryotic host cells	p. 235
Adding tags and signals	p. 239
In vitro mutagenesis	p. 242
Vaccines	p. 247
Genomic analysis	p. 251
Genome sequencing	p. 251
Analysis and annotation	p. 261
Comparing genomes	p. 269
Genome browsers	p. 271
Relating genes and functions: genetic and physical maps	p. 273
Transposon mutagenesis and other screening techniques	p. 276
Conclusion	p. 285
Analysis of genetic variation	p. 287
Single nucleotide polymorphisms	p. 288
Larger-scale variations	p. 291
Other methods for studying variation	p. 293
Human genetic diseases	p. 300
Molecular phylogeny	p. 305
Post-genomic analysis	p. 313
Analysing transcription; transcriptomes	p. 313
Array-based methods	p. 319
Translational analysis; proteomics	p. 326
Post-translational analysis: protein interactions	p. 329
Integrative studies; systems biology	p. 332
Modifying organisms; transgenics	p. 333
Modification of bacteria and viruses: live vaccines	p. 333
Transgenesis and cloning	p. 337
Animal transgenesis	p. 338
Applications of transgenic animals	p. 347
Transgenic plants and their applications	p. 351
Glossary	p. 355
Bibliography	p. 373
Index	p. 375
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