An Introduction to Genetic EngineeringDes Nicholl presents here a new, fully revised, and expanded edition of his popular undergraduate-level textbook. Many of the features of the original edition have been retained; the book still offers a concise technical introduction to the subject of genetic engineering. However, the book is now divided into three main sections: the first introduces students to basic molecular biology, the second section explains the methods used to manipulate genes, and the third deals with modern applications of genetic engineering. A whole chapter is now devoted to the polymerase chain reaction. Applications covered in the book include genomics, protein engineering, gene therapy, cloning, and transgenic animals and plants. A final chapter discusses the ethical questions surrounding genetic engineering in general. An Introduction to Genetic Engineering is essential reading for undergraduate students of biotechnology, genetics, molecular biology and biochemistry. |
Contents
Introduction | 1 |
12 Laying the foundations | 3 |
13 First steps | 4 |
14 What is in store? | 6 |
The basis of genetic engineering | 9 |
Introducing molecular biology | 11 |
22 The structure of DNA and RNA | 13 |
23 Gene organisation | 16 |
75 Applications of the PCR | 130 |
Selection screening and analysis of recombinants | 132 |
81 Genetic selection and screening methods | 133 |
82 Screening using nucleic acid hybridisation | 138 |
83 Immunological screening for expressed genes | 141 |
84 Analysis of cloned genes | 143 |
Genetic engineering in action | 151 |
Understanding genes and genomes | 153 |
24 Gene expression | 21 |
25 Genes and genomes | 23 |
Working with nucleic acids | 27 |
32 Handling and quantification of nucleic acids | 29 |
33 Radiolabelling of nucleic acids | 30 |
34 Nucleic acid hybridisation | 33 |
36 DNA sequencing | 35 |
The tools of the trade | 43 |
42 DNA modifying enzymes | 48 |
43 DNA ligase joining DNA molecules | 52 |
The methodology of gene manipulation | 55 |
Host cells and vectors | 57 |
51 Host cell types | 58 |
52 Plasmid vectors for use in E coli | 60 |
53 Bacteriophage vectors for use in E coli | 66 |
54 Other vectors | 75 |
55 Getting DNA into cells | 80 |
Cloning strategies | 87 |
62 Cloning from mRNA | 89 |
63 Cloning from genomic DNA | 98 |
64 Advanced cloning strategies | 106 |
The polymerase chain reaction | 115 |
72 The methodology of the PCR | 118 |
73 More exotic PCR techniques | 123 |
74 Processing of PCR products | 129 |
92 From genes to genomes | 159 |
93 Genome sequencing | 165 |
94 The human genome project | 167 |
Genetic engineering and biotechnology | 178 |
101 Making proteins | 179 |
102 Protein engineering | 183 |
103 Examples of biotechnological applications of rDNA technology | 185 |
Medical and forensic applications of gene manipulation | 197 |
112 Treatment using rDNA technology gene therapy | 210 |
113 DNA profiling | 215 |
Transgenic plants and animals | 224 |
122 Transgenic animals | 237 |
The other sort of cloning | 247 |
132 Frogs and toads and carrots | 250 |
133 A famous sheep the breakthrough achieved | 253 |
134 Beyond Dolly | 256 |
Brave new world or genetic nightmare? | 259 |
142 Elements of the ethics debate | 260 |
141 Does Frankensteins monster live inside Pandoras box? | 262 |
| 263 | |
Using the World Wide Web | 266 |
Glossary | 270 |
| 287 | |
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Common terms and phrases
allele amino acid amplified analysis applications B-galactosidase bacterial bacteriophage BamHI base-pairs biotechnology cDNA Chapter chromosome cloned DNA cloned sequences coli complex copy defective digestion disease DNA fragments DNA ligase DNA molecules DNA polymerase DNA profiling DNA sequence double-stranded EcoRI embryo enable encodes eukaryotic exonuclease function gene cloning gene expression gene manipulation gene sequence gene therapy genetic engineering genomic DNA genomic library host cell human genome hybrid hybridisation infection involved isolated known labelled lacZ ligation markers methods molecular biology mouse mRNA mutation nuclear nucleic acid nucleotide oligonucleotide organism phage phenotype physical mapping plaques plasmid vectors primer probe problems produced protein reaction recombinant DNA region replacement Reproduced with permission restriction enzyme restriction map screening selection shown in Fig single-stranded specific strand synthesis target technique tion transcription transgenic animals transgenic plants translation types usually vitro VNTR yeast