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9780521004718

An Introduction to Genetic Engineering

by
  • ISBN13:

    9780521004718

  • ISBN10:

    0521004713

  • Edition: 2nd
  • Format: Paperback
  • Copyright: 2002-02-11
  • Publisher: Cambridge University Press
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List Price: $44.99

Summary

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.

Author Biography

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

Table of Contents

Preface to the second edition xi
Introduction
1(8)
What is genetic engineering?
1(2)
Laying the foundations
3(1)
First steps
4(2)
What is in store?
6(3)
Part I: The basis of genetic engineering 9(46)
Introducting molecular biology
11(16)
The flow of genetic information
11(2)
The structure of DNA and RNA
13(3)
Gene organisation
16(5)
Gene structure in prokaryotes
18(1)
Gene structure in eukaryotes
19(2)
Gene expression
21(2)
Genes and genomes
23(4)
Genome size and complexity
23(1)
Genome organisation
24(3)
Working with nucleic acids
27(16)
Isolation of DNA and RNA
27(2)
Handling and quantification of nucleic acids
29(1)
Radiolabelling of nucleic acids
30(3)
End labelling
30(1)
Nick translation
31(1)
Labelling by primer extension
31(2)
Nucleic acid hybridisation
33(1)
Gel electrophoresis
33(2)
DNA sequencing
35(8)
Maxam-Gilbert (chemical) sequencing
37(1)
Sanger-Coulson (dideoxy or enzymatic) sequencing
37(3)
Electrophoresis and reading of sequences
40(3)
The tools of the trade
43(12)
Restriction enzymes - cutting DNA
43(5)
Type II restriction endonucleases
44(1)
Use of restriction endonucleases
45(2)
Restriction mapping
47(1)
DNA modifying enzymes
48(4)
Nucleases
48(1)
Polymerases
49(2)
Enzymes that modify the ends of DNA molecules
51(1)
DNA ligase - joining DNA molecules
52(3)
Part II: The methodology of gene manipulation 55(96)
Host cells and vectors
57(30)
Host cell types
58(2)
Prokaryotic hosts
58(1)
Eukaryotic hosts
59(1)
Plasmid vectors for use in E. coli
60(6)
What are plasmids?
61(1)
Basic cloning plasmids
61(2)
Slightly more exotic plasmid vectors
63(3)
Bacteriophage vectors for use in E. coli
66(9)
What are bacteriophages?
66(4)
Vectors based on bacteriophage λ
70(4)
Vectors based on bacteriophage M13
74(1)
Other vectors
75(5)
Hybrid plasmid/phage vectors
76(1)
Vectors for use in eukaryotic cells
77(2)
Artificial chromosomes
79(1)
Getting DNA into cells
80(7)
Transformation and transfection
80(1)
Packaging phage DNA in vitro
81(2)
Alternative DNA delivery methods
83(4)
Cloning strategies
87(28)
Which approach is best?
87(2)
Cloning from mRNA
89(9)
Synthesis of cDNA
90(3)
Cloning cDNA in plasmid vectors
93(3)
Cloning cDNA in bacteriophage vectors
96(2)
Cloning from genomic DNA
98(8)
Genomic libraries
99(2)
Preparation of DNA fragments for cloning
101(2)
Ligation, packaging and amplification of libraries
103(3)
Advanced cloning strategies
106(9)
Synthesis and cloning of cDNA
106(3)
Expression of cloned cDNA molecules
109(2)
Cloning large DNA fragments in BAC and YAC vectors
111(4)
The polymerase chain reaction
115(17)
The (short) history of the PCR
115(3)
The methodology of the PCR
118(5)
The essential features of the PCR
118(3)
The design of primers for PCR
121(1)
DNA polymerases for PCR
121(2)
More exotic PCR techniques
123(6)
PCR using mRNA templates
123(1)
Nested PCR
124(2)
Inverse PCR
126(1)
RAPD and several other acronyms
127(2)
Processing of PCR products
129(1)
Applications of the PCR
130(2)
Selection, screening and analysis of recombinants
132(19)
Genetic selection and screening methods
133(5)
The use of chromogenic substrates
133(2)
Insertional inactivation
135(1)
Complementation of defined mutations
136(1)
Other genetic selection methods
137(1)
Screening using nucleic acid hybridisation
138(3)
Nucleic acid probes
138(1)
Screening clone banks
139(2)
Immunological screening for expressed genes
141(2)
Analysis of cloned genes
143(8)
Characterisation based on mRNA translation in vitro
143(2)
Restriction mapping
145(1)
Blotting techniques
145(3)
DNA sequencing
148(3)
Part III: Genetic engineering in action 151(112)
Understanding genes and genomes
153(25)
Analysis of gene structure and function
153(6)
A closer look at sequences
154(1)
Finding important regions of genes
155(2)
Investigating gene expression
157(2)
From genes to genomes
159(6)
Analysing genomes
160(1)
Mapping genomes
161(4)
Genome sequencing
165(2)
Sequencing technology
165(1)
Genome projects
165(2)
The human genome project
167(11)
Whose genome, and how many genes does it contain?
169(1)
Genetic and physical maps of the human genome
170(4)
Deriving and assembling the sequence
174(1)
What next?
175(3)
Genetic engineering and biotechnology
178(19)
Making proteins
179(4)
Native and fusion proteins
179(2)
Yeast expression systems
181(1)
The baculovirus expression system
182(1)
Mammalian cell lines
183(1)
Protein engineering
183(2)
Examples of biotechnological applications of rDNA technology
185(12)
Production of enzymes
185(2)
The BST story
187(3)
Therapeutic products for use in human health-care
190(7)
Medical and forensic applications of gene manipulation
197(27)
Diagnosis and characterisation of medical conditions
197(13)
Diagnosis of infection
198(1)
Patterns of inheritance
198(3)
Genetically based disease conditions
201(9)
Treatment using rDNA technology - gene therapy
210(5)
Getting transgenes into patients
211(3)
Gene therapy for adenosine deaminase deficiency
214(1)
Gene therapy for cystic fibrosis
214(1)
DNA profiling
215(9)
The history of `genetic fingerprinting'
216(2)
DNA profiling and the law
218(1)
Mysteries of the past revealed by genetic detectives
219(5)
Transgenic plants and animals
224(23)
Transgenic plants
224(13)
Why transgenic plants?
225(1)
Tiplasmids as vectors for plant cells
226(2)
Making transgenic plants
228(2)
Putting the technology to work
230(7)
Transgenic animals
237(10)
Why transgenic animals?
237(1)
Producting transgenic animals
238(3)
Applications of transgenic animal technology
241(6)
The other sort of cloning
247(12)
Early thoughts and experiments
247(3)
First steps towards cloning
249(1)
Nuclear totipotency
250(1)
Frogs and toads and carrots
250(3)
A famous sheep - the breakthrough achieved
253(3)
Beyond Dolly
256(3)
Brave new world or genetic nightmare?
259(4)
Is science ethically and morally neutral?
259(1)
Elements of the ethics debate
260(2)
Does Frankenstein's monster live inside Pandora's box?
262(1)
Suggestings for further reading 263(3)
Using the World Wide Web 266(4)
Glossary 270(17)
Index 287

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