An Introduction to Genetic Engineering / Desmond S.T. Nicholl.

By:

Nicholl, Desmond S. T

Material type: Text

TextSeries: Studies in BiologyPublication details: Cambridge: Cambridge University Press, c.2002.Edition: 2nd edDescription: xii, 292 p. : ill. ; 24 cmISBN:

0521004713 (pbk.)

Subject(s):

Genetic engineering

DDC classification:

660.65 NIC

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Standard Loan	Moylish Library Main Collection	660.65 NIC (Browse shelf(Opens below))	1	Available		39002000388349

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Enhanced descriptions from Syndetics:

Des 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.

Includes bibliographical references and index.

Table of contents provided by Syndetics

Preface to the second edition (p. xi)
1 Introduction (p. 1)
1.1 What is genetic engineering? (p. 1)
1.2 Laying the foundations (p. 3)
1.3 First steps (p. 4)
1.4 What is in store? (p. 6)
Part I The basis of genetic engineering (p. 9)
2 Introducing molecular biology (p. 11)
2.1 The flow of genetic information (p. 11)
2.2 The structure of DNA and RNA (p. 13)
2.3 Gene organisation (p. 16)
2.3.1 Gene structure in prokaryotes (p. 18)
2.3.2 Gene structure in eukaryotes (p. 19)
2.4 Gene expression (p. 21)
2.5 Genes and genomes (p. 23)
2.5.1 Genome size and complexity (p. 23)
2.5.2 Genome organisation (p. 24)
3 Working with nucleic acids (p. 27)
3.1 Isolation of DNA and RNA (p. 27)
3.2 Handling and quantification of nucleic acids (p. 29)
3.3 Radiolabelling of nucleic acids (p. 30)
3.3.1 End labelling (p. 30)
3.3.2 Nick translation (p. 31)
3.3.3 Labelling by primer extension (p. 31)
3.4 Nucleic acid hybridisation (p. 33)
3.5 Gel electrophoresis (p. 33)
3.6 DNA sequencing (p. 35)
3.6.1 Maxam--Gilbert (chemical) sequencing (p. 37)
3.6.2 Sanger--Coulson (dideoxy or enzymatic) sequencing (p. 37)
3.6.3 Electrophoresis and reading of sequences (p. 40)
4 The tools of the trade (p. 43)
4.1 Restriction enzymes--cutting DNA (p. 43)
4.1.1 Type II restriction endonucleases (p. 44)
4.1.2 Use of restriction endonucleases (p. 45)
4.1.3 Restriction mapping (p. 47)
4.2 DNA modifying enzymes (p. 48)
4.2.1 Nucleases (p. 48)
4.2.2 Polymerases (p. 49)
4.2.3 Enzymes that modify the ends of DNA molecules (p. 51)
4.3 DNA ligase--joining DNA molecules (p. 52)
Part II The methodology of gene manipulation (p. 55)
5 Host cells and vectors (p. 57)
5.1 Host cell types (p. 58)
5.1.1 Prokaryotic hosts (p. 58)
5.1.2 Eukaryotic hosts (p. 59)
5.2 Plasmid vectors for use in E. coli (p. 60)
5.2.1 What are plasmids? (p. 61)
5.2.2 Basic cloning plasmids (p. 61)
5.2.3 Slightly more exotic plasmid vectors (p. 63)
5.3 Bacteriophage vectors for use in E. coli (p. 66)
5.3.1 What are bacteriophages? (p. 66)
5.3.2 Vectors based on bacteriophage [lambda] (p. 70)
5.3.3 Vectors based on bacteriophage M13 (p. 74)
5.4 Other vectors (p. 75)
5.4.1 Hybrid plasmid/phage rectors (p. 76)
5.4.2 Vectors for use in eukaryotic cells (p. 77)
5.4.3 Artificial chromosomes (p. 79)
5.5 Getting DNA into cells (p. 80)
5.5.1 Transformation and transfection (p. 80)
5.5.2 Packaging phage DNA in vitro (p. 81)
5.5.3 Alternative DNA delivery methods (p. 83)
6 Cloning strategies (p. 87)
6.1 Which approach is best? (p. 87)
6.2 Cloning from mRNA (p. 89)
6.2.1 Synthesis of cDNA (p. 90)
6.2.2 Cloning cDNA in plasmid vectors (p. 93)
6.2.3 Cloning cDNA in bacteriophage vectors (p. 96)
6.3 Cloning from genomic DNA (p. 98)
6.3.1 Genomic libraries (p. 99)
6.3.2 Preparation of DNA fragments for cloning (p. 101)
6.3.3 Ligation, packaging and amplification of libraries (p. 103)
6.4 Advanced cloning strategies (p. 106)
6.4.1 Synthesis and cloning of cDNA (p. 106)
6.4.2 Expression of cloned cDNA molecules (p. 109)
6.4.3 Cloning large DNA fragments in BAC and YAC vectors (p. 111)
7 The polymerase chain reaction (p. 115)
7.1 The (short) history of the PCR (p. 115)
7.2 The methodology of the PCR (p. 118)
7.2.1 The essential features of the PCR (p. 118)
7.2.2 The design of primers for PCR (p. 121)
7.2.3 DNA polymerases for PCR (p. 121)
7.3 More exotic PCR techniques (p. 123)
7.3.1 PCR using mRNA templates (p. 123)
7.3.2 Nested PCR (p. 124)
7.3.3 Inverse PCR (p. 126)
7.3.4 RAPD and several other acronyms (p. 127)
7.4 Processing of PCR products (p. 129)
7.5 Applications of the PCR (p. 130)
8 Selection, screening and analysis of recombinants (p. 132)
8.1 Genetic selection and screening methods (p. 133)
8.1.1 The use of chromogenic substrates (p. 133)
8.1.2 Insertional inactivation (p. 135)
8.1.3 Complementation of defined mutations (p. 136)
8.1.4 Other genetic selection methods (p. 137)
8.2 Screening using nucleic acid hybridisation (p. 138)
8.2.1 Nucleic acid probes (p. 138)
8.2.2 Screening clone banks (p. 139)
8.3 Immunological screening for expressed genes (p. 141)
8.4 Analysis of cloned genes (p. 143)
8.4.1 Characterisation based on mRNA translation in vitro (p. 143)
8.4.2 Restriction mapping (p. 145)
8.4.3 Blotting techniques (p. 145)
8.4.4 DNA sequencing (p. 148)
Part III Genetic engineering in action (p. 151)
9 Understanding genes and genomes (p. 153)
9.1 Analysis of gene structure and function (p. 153)
9.1.1 A closer look at sequences (p. 154)
9.1.2 Finding important regions of genes (p. 155)
9.1.3 Investigating gene expression (p. 157)
9.2 From genes to genomes (p. 159)
9.2.1 Analysing genomes (p. 160)
9.2.2 Mapping genomes (p. 161)
9.3 Genome sequencing (p. 165)
9.3.1 Sequencing technology (p. 165)
9.3.2 Genome projects (p. 165)
9.4 The human genome project (p. 167)
9.4.1 Whose genome, and how many genes does it contain? (p. 169)
9.4.2 Genetic and physical maps of the human genome (p. 170)
9.4.3 Deriving and assembling the sequence (p. 174)
9.4.4 What next? (p. 175)
10 Genetic engineering and biotechnology (p. 178)
10.1 Making proteins (p. 179)
10.1.1 Native and fusion proteins (p. 179)
10.1.2 Yeast expression systems (p. 181)
10.1.3 The baculovirus expression system (p. 182)
10.1.4 Mammalian cell lines (p. 183)
10.2 Protein engineering (p. 183)
10.3 Examples of biotechnological applications of rDNA technology (p. 185)
10.3.1 Production of enzymes (p. 185)
10.3.2 The BST story (p. 187)
10.3.3 Therapeutic products for use in human health-care (p. 190)
11 Medical and forensic applications of gene manipulation (p. 197)
11.1 Diagnosis and characterisation of medical conditions (p. 197)
11.1.1 Diagnosis of infection (p. 198)
11.1.2 Patterns of inheritance (p. 198)
11.1.3 Genetically based disease conditions (p. 201)
11.2 Treatment using rDNA technology--gene therapy (p. 210)
11.2.1 Getting transgenes into patients (p. 211)
11.2.2 Gene therapy for adenosine deaminase deficiency (p. 214)
11.2.3 Gene therapy for cystic fibrosis (p. 214)
11.3 DNA profiling (p. 215)
11.3.1 The history of 'genetic fingerprinting' (p. 216)
11.3.2 DNA profiling and the law (p. 218)
11.3.3 Mysteries of the past revealed by genetic detectives (p. 219)
12 Transgenic plants and animals (p. 224)
12.1 Transgenic plants (p. 224)
12.1.1 Why transgenic plants? (p. 225)
12.1.2 Ti plasmids as vectors for plant cells (p. 226)
12.1.3 Making transgenic plants (p. 228)
12.1.4 Putting the technology to work (p. 230)
12.2 Transgenic animals (p. 237)
12.2.1 Why transgenic animals? (p. 237)
12.2.2 Producing transgenic animals (p. 238)
12.2.3 Applications of transgenic animal technology (p. 241)
13 The other sort of cloning (p. 247)
13.1 Early thoughts and experiments (p. 247)
13.1.1 First steps towards cloning (p. 249)
13.1.2 Nuclear totipotency (p. 250)
13.2 Frogs and toads and carrots (p. 250)
13.3 A famous sheep--the breakthrough achieved (p. 253)
13.4 Beyond Dolly (p. 256)
14 Brave new world or genetic nightmare? (p. 259)
14.1 Is science ethically and morally neutral? (p. 259)
14.2 Elements of the ethics debate (p. 260)
14.1 Does Frankenstein's monster live inside Pandora's box? (p. 262)
Suggestions for further reading (p. 263)
Using the World Wide Web (p. 266)
Glossary (p. 270)
Index (p. 287)

Author notes provided by Syndetics

Des Nicholl is a Senior Lecturer in Biological Sciences, The University of Paisley, Scotland, UK