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[资源] Animal Transgenesis and Cloning

1 From the gene to the transgenic animal 1
1.1 Genome composition 1
1.2 Gene structure 4
1.3 The number of genes in genomes 7
1.4 The major techniques of genetic engineering 13
1.4.1 Gene cloning 13
1.4.2 DNA sequencing 14
1.4.3 In vitro gene amplification 14
1.4.4 Gene construction 14
1.4.5 Gene transfer into cells 16
1.5 The systematic description of genomes 21
1.6 Classical genetic selection 26
1.7 Experimental mutation in genomes 27
1.7.1 Chemical mutagenesis 27
1.7.2 Mutagenesis by integration of foreign DNA 29
1.7.3 Mutagenesis by transgenesis 30
2 Techniques for cloning and transgenesis 33
2.1 Cloning 33
2.1.1 The main steps of differentiation 33
2.1.2 Cloning by nuclear transfer 37
2.2 Gene therapy 48
2.2.1 The goals of gene therapy 48
2.2.2 The tools of gene therapy 49
2.2.3 The applications of gene therapy 52
2.3 Techniques of animal transgenesis 54
2.3.1 The aims and the concept of animal transgenesis 54
2.3.2 Gene transfer into gametes 60
2.3.3 Gene transfer into embryos 65
2.3.4 Gene transfer via cells 69
2.3.5 Vectors for gene addition 73
2.3.6 Vectors for gene replacement 85
2.3.7 Vectors for the rearrangement of
targeted genes 90
2.3.8 Targeted integration of foreign genes 97
2.3.9 Non-classical vectors for the recombination
of targeted genes 105
2.3.10 Vectors for gene trap 106
2.3.11 Vectors for the expression of transgenes 116
3 Applications of cloning and transgenesis 137
3.1 Applications of animal cloning 137
3.1.1 Basic research 137
3.1.2 Transgenesis 142
3.1.3 Animal reproduction 143
3.1.4 Human reproduction 144
3.1.5 Therapeutic cloning 144
3.1.6 Xenografting 150
3.2 Applications of animal transgenesis 153
3.2.1 Basic research 153
3.2.2 Study of human diseases 154
3.2.3 Pharmaceutical production 159
3.2.4 Xenografting 162
3.2.5 Breeding 163
4 Limits and risks of cloning, gene therapy
and transgenesis 171
4.1 Limits and risks of cloning 173
4.1.1 Reproductive cloning in humans 173
4.1.2 Reproductive cloning in animals 175
4.1.3 Therapeutic cloning 176
4.2 Limits and risks of gene therapy 177
4.3 Limits and risks of transgenesis 178
4.3.1 Technical and theoretical limits 178
4.3.2 Biosafety problems in confined areas 179
4.3.3 The intentional dissemination of transgenic
animals into the environment 181
4.3.4 The risks for human consumers 184
4.3.5 Transgenesis and animal welfare 185
vi CONTENTS
4.3.6 Patenting of transgenic animals 187
4.3.7 Transgenesis in humans 188
Conclusion and Perspectives 191
References 199
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