From genes to genomes: Concepts and applications of DNA technology

✍ Scribed by Dale J., von Schantz M.

• Publisher: Wiley
• Year: 2007
• Tongue: English
• Leaves: 396
• Edition: 2ed
• Category: Library

✦ Synopsis

“… an excellent book… achieves all of its goals with style, clarity and completeness… You can see the power and possibilities of molecular genetics as you read…” –Human Genetics"This volume hits an outstanding balance among readability, coverage, and detail." –Biochemistry and Molecular Biology EducationRapid 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 themeUpdating and reinforcing of material on proteomics, gene therapy and stem cellsMore eukaryotic/mammalian examples and less focus on bacteriaThis 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.

✦ Table of Contents

From Genes to Genomes......Page 5
Contents......Page 7
Preface......Page 11
1: Introduction......Page 13
2.1 Nucleic acid structure......Page 19
2.2 What is a gene?......Page 27
2.3 Information flow: gene expression......Page 28
2.4 Gene structure and organization......Page 33
3.1 What is cloning?......Page 37
3.2 Overview of the procedures......Page 38
3.3 Gene libraries......Page 41
3.4 Hybridization......Page 42
3.5 Polymerase chain reaction......Page 44
3.6 Extraction and purification of nucleic acids......Page 45
3.7 Detection and quantitation of nucleic acids......Page 48
3.8 Gel electrophoresis......Page 49
4.1 Restriction endonucleases......Page 53
4.2 Ligation......Page 59
4.3 Modification of restriction fragment ends......Page 65
4.4 Other ways of joining DNA molecules......Page 69
5.1 Plasmid vectors......Page 73
5.2 Vectors based on the lambda bacteriophage......Page 81
5.3 Cosmids......Page 90
5.4 M13 vectors......Page 91
5.5 Expression vectors......Page 93
5.6 Vectors for cloning and expression in eukaryotic cells......Page 96
5.7 Supervectors: YACs and BACs......Page 104
5.8 Summary......Page 106
6: Genomic and cDNA Libraries......Page 107
6.1 Genomic libraries......Page 108
6.2 Growing and storing libraries......Page 117
6.3 cDNA libraries......Page 118
6.4 Random, arrayed and ordered libraries......Page 125
7.1 Screening libraries with gene probes......Page 129
7.2 Screening expression libraries with antibodies......Page 140
7.3 Subcloning......Page 142
7.4 Characterization of plasmid clones......Page 143
8: Polymerase Chain Reaction......Page 147
8.1 The PCR reaction......Page 148
8.2 PCR in practice......Page 152
8.3 Cloning PCR products......Page 156
8.5 Reverse-transcription PCR......Page 158
8.6 Rapid amplification of cDNA ends......Page 159
8.7 Quantitative PCR......Page 161
8.8 Applications of PCR......Page 166
9.1 DNA sequencing......Page 171
9.2 Databank entries and annotation......Page 179
9.3 Sequence analysis......Page 185
9.4 Sequence comparisons......Page 190
9.5 Protein structure......Page 202
9.6 Confirming gene function......Page 208
10.1 Analysing transcription......Page 213
10.2 Methods for studying the promoter......Page 221
10.3 Regulatory elements and DNA-binding proteins......Page 225
10.4 Translational analysis......Page 229
11: Products from Native and Manipulated Cloned Genes......Page 233
11.1 Factors affecting expression of cloned genes......Page 234
11.2 Expression of cloned genes in bacteria......Page 239
11.3 Expression in eukaryotic host cells......Page 247
11.4 Adding tags and signals......Page 251
11.5 In vitro mutagenesis......Page 254
11.6 Vaccines......Page 259
12.1 Genome sequencing......Page 263
12.2 Analysis and annotation......Page 273
12.3 Comparing genomes......Page 281
12.4 Genome browsers......Page 283
12.5 Relating genes and functions: genetic and physical maps......Page 285
12.6 Transposon mutagenesis and other screening techniques......Page 288
12.7 Conclusion......Page 297
13: Analysis of Genetic Variation......Page 299
13.1 Single nucleotide polymorphisms......Page 300
13.2 Larger-scale variations......Page 303
13.3 Other methods for studying variation......Page 305
13.4 Human genetic diseases......Page 312
13.5 Molecular phylogeny......Page 317
14.1 Analysing transcription; transcriptomes......Page 325
14.2 Array-based methods......Page 331
14.3 Translational analysis; proteomics......Page 338
14.4 Post-translational analysis: protein interactions......Page 341
14.5 Integrative studies; systems biology......Page 344
15.1 Modification of bacteria and viruses: live vaccines......Page 345
15.2 Transgenesis and cloning......Page 349
15.3 Animal transgenesis......Page 350
15.4 Applications of transgenic animals......Page 359
15.5 Transgenic plants and their applications......Page 363
Glossary......Page 367
Bibliography......Page 385
Index......Page 387