Introduction to Genetic Engineering
Third Edition
In this third edition of his popular undergraduate-level textbook, Desmond Nicholl
recognises that a sound grasp of basic principles is vital in any introduction to
genetic engineering. Therefore, as well as being thoroughly updated, the book also
retains its focus on the fundamental principles used in gene manipulation. The text
is divided into three sections: Part I provides an introduction to the relevant basic
molecular biology; Part II, the methods used to manipulate genes; and Part III,
applications of the technology. There is a new chapter devoted to the emerging
importance of bioinformatics as a distinct discipline. Other additional features
include text boxes, which highlight important aspects of topics discussed, and
chapter summaries, which include aims and learning outcomes. These, along with
key word listings, concept maps, and a glossary, will enable students to tailor their
studies to suit their own learning styles and ultimately gain a firm grasp on this
subject that students traditionally find difficult.
Desmond S. T. Nicholl is a Senior Lecturer in Biological Sciences at the University
of the West of Scotland, Paisley, UK.
, Desmond S. T. Nicholl
University of the West of Scotland, Paisley, UK
An Introduction to
Genetic Engineering
ThirdEdition
,CAMBRIDGE UNIVERSITY PRESS
3.5 Nucleic acid hybridisation 39
Cambridge, New York, Melbourne, Madrid, Cape Town, Singapore, São Paulo
Contents
Cambridge University Press
The Edinburgh Building, Cambridge CB2 8RU, UK
Published in the United States of America by Cambridge University Press, New Preface to the third edition page xi
York www.cambridge.org Chapter 1 Introduction 2
Information on this title: www.cambridge.org/9780521850063 Chapter summary 2
1.1 What is genetic engineering? 3
© Desmond S. T. Nicholl 2008
1.2 Laying the foundations 5
1.3 First steps 6
This publication is in copyright. Subject to statutory exception and to the provision 1.4 What’s in store? 7
of relevant collective licensing agreements, no reproduction of any part may take Concept map 10
place without the written permission of Cambridge University Press.
Part I The basis of genetic engineering
First published in print format 2008
Chapter 2 Introducing molecular biology 12
Chapter summary 12
ISBN-13 978-0-511-39858-2 eBook (EBL)
2.1 The way that living systems are organised 13
2.2 The flow of genetic information 15
ISBN-13 978-0-521-85006-3 hardback
2.3 The structure of DNA and RNA 16
ISBN-13 978-0-521-61521-1 paperback 2.4 Gene organisation 19
2.4.1 The anatomy of a gene 20
2.4.2 Gene structure in prokaryotes 21
2.4.3 Gene structure in eukaryotes 22
2.5 Gene expression 23
Cambridge University Press has no responsibility for the persistence or accuracy of
2.5.1 From genes to proteins 24
urls for external or third-party internet websites referred to in this publication, and
2.5.2 Transcription and translation 24
does not guarantee that any content on such websites is, or will remain, accurate or
2.5.3 Regulation of gene expression 25
appropriate. 2.6 Genes and genomes 27
2.6.1 Genome size and complexity 28
Chapter 3 Working with nucleic acids 32 2.6.2 Genome organisation 29
Chapter summary 32 2.6.3 The transcriptome and proteome 30
3.1 Laboratory requirements 33 Concept map 31
3.2 Isolation of DNA and RNA 34
3.3 Handling and quantification of nucleic acids 35
3.4 Labelling nucleic acids 37
3.4.1 Types of label -- radioactive or not? 37
3.4.2 End labelling 38
3.4.3 Nick translation 38
3.4.4 Labelling by primer extension 39
, vi
3.6 Gel electrophoresis 40
3.7 DNA sequencing 41
3.7.1 Principles of DNA sequencing 42
3.7.2 Preparation of DNA fragments 43
3.7.3 Maxam--Gilbert (chemical) sequencing 44
3.7.4 Sanger--Coulson (dideoxy or enzymatic) sequencing 45
3.7.5 Electrophoresis and reading of sequences 47
3.7.6 Automation of DNA sequencing 48
Concept map 49
Chapter 4 The tools of the trade 50
Chapter summary 50
4.1 Restriction enzymes -- cutting DNA 51
4.1.1 Type II restriction endonucleases 52
4.1.2 Use of restriction endonucleases 53
4.1.3 Restriction mapping 55
4.2 DNA modifying enzymes 55
4.2.1 Nucleases 56
4.2.2 Polymerases 57
4.2.3 Enzymes that modify the ends of DNA molecules 58
4.3 DNA ligase -- joining DNA molecules 58
Concept map 60
Chapter 5 Host cells and vectors 62
Chapter summary 62
5.1 Host cell types 64
5.1.1 Prokaryotic hosts 64
5.1.2 Eukaryotic hosts 65
5.2 Plasmid vectors for use in E. coli 66
5.2.1 What are plasmids? 66
5.2.2 Basic cloning plasmids 67
5.2.3 Slightly more exotic plasmid vectors 69
5.3 Bacteriophage vectors for use in E. coli 70
5.3.1 What are bacteriophages? 71
5.3.2 Vectors based on bacteriophage 75
5.3.3 Vectors based on bacteriophage M13 78
5.4 Other vectors 79
5.4.1 Hybrid plasmid/phage vectors 80
5.4.2 Vectors for use in eukaryotic cells 80
5.4.3 Artificial chromosomes 83
5.5 Getting DNA into cells 84
5.5.1 Transformation and transfection 84
Third Edition
In this third edition of his popular undergraduate-level textbook, Desmond Nicholl
recognises that a sound grasp of basic principles is vital in any introduction to
genetic engineering. Therefore, as well as being thoroughly updated, the book also
retains its focus on the fundamental principles used in gene manipulation. The text
is divided into three sections: Part I provides an introduction to the relevant basic
molecular biology; Part II, the methods used to manipulate genes; and Part III,
applications of the technology. There is a new chapter devoted to the emerging
importance of bioinformatics as a distinct discipline. Other additional features
include text boxes, which highlight important aspects of topics discussed, and
chapter summaries, which include aims and learning outcomes. These, along with
key word listings, concept maps, and a glossary, will enable students to tailor their
studies to suit their own learning styles and ultimately gain a firm grasp on this
subject that students traditionally find difficult.
Desmond S. T. Nicholl is a Senior Lecturer in Biological Sciences at the University
of the West of Scotland, Paisley, UK.
, Desmond S. T. Nicholl
University of the West of Scotland, Paisley, UK
An Introduction to
Genetic Engineering
ThirdEdition
,CAMBRIDGE UNIVERSITY PRESS
3.5 Nucleic acid hybridisation 39
Cambridge, New York, Melbourne, Madrid, Cape Town, Singapore, São Paulo
Contents
Cambridge University Press
The Edinburgh Building, Cambridge CB2 8RU, UK
Published in the United States of America by Cambridge University Press, New Preface to the third edition page xi
York www.cambridge.org Chapter 1 Introduction 2
Information on this title: www.cambridge.org/9780521850063 Chapter summary 2
1.1 What is genetic engineering? 3
© Desmond S. T. Nicholl 2008
1.2 Laying the foundations 5
1.3 First steps 6
This publication is in copyright. Subject to statutory exception and to the provision 1.4 What’s in store? 7
of relevant collective licensing agreements, no reproduction of any part may take Concept map 10
place without the written permission of Cambridge University Press.
Part I The basis of genetic engineering
First published in print format 2008
Chapter 2 Introducing molecular biology 12
Chapter summary 12
ISBN-13 978-0-511-39858-2 eBook (EBL)
2.1 The way that living systems are organised 13
2.2 The flow of genetic information 15
ISBN-13 978-0-521-85006-3 hardback
2.3 The structure of DNA and RNA 16
ISBN-13 978-0-521-61521-1 paperback 2.4 Gene organisation 19
2.4.1 The anatomy of a gene 20
2.4.2 Gene structure in prokaryotes 21
2.4.3 Gene structure in eukaryotes 22
2.5 Gene expression 23
Cambridge University Press has no responsibility for the persistence or accuracy of
2.5.1 From genes to proteins 24
urls for external or third-party internet websites referred to in this publication, and
2.5.2 Transcription and translation 24
does not guarantee that any content on such websites is, or will remain, accurate or
2.5.3 Regulation of gene expression 25
appropriate. 2.6 Genes and genomes 27
2.6.1 Genome size and complexity 28
Chapter 3 Working with nucleic acids 32 2.6.2 Genome organisation 29
Chapter summary 32 2.6.3 The transcriptome and proteome 30
3.1 Laboratory requirements 33 Concept map 31
3.2 Isolation of DNA and RNA 34
3.3 Handling and quantification of nucleic acids 35
3.4 Labelling nucleic acids 37
3.4.1 Types of label -- radioactive or not? 37
3.4.2 End labelling 38
3.4.3 Nick translation 38
3.4.4 Labelling by primer extension 39
, vi
3.6 Gel electrophoresis 40
3.7 DNA sequencing 41
3.7.1 Principles of DNA sequencing 42
3.7.2 Preparation of DNA fragments 43
3.7.3 Maxam--Gilbert (chemical) sequencing 44
3.7.4 Sanger--Coulson (dideoxy or enzymatic) sequencing 45
3.7.5 Electrophoresis and reading of sequences 47
3.7.6 Automation of DNA sequencing 48
Concept map 49
Chapter 4 The tools of the trade 50
Chapter summary 50
4.1 Restriction enzymes -- cutting DNA 51
4.1.1 Type II restriction endonucleases 52
4.1.2 Use of restriction endonucleases 53
4.1.3 Restriction mapping 55
4.2 DNA modifying enzymes 55
4.2.1 Nucleases 56
4.2.2 Polymerases 57
4.2.3 Enzymes that modify the ends of DNA molecules 58
4.3 DNA ligase -- joining DNA molecules 58
Concept map 60
Chapter 5 Host cells and vectors 62
Chapter summary 62
5.1 Host cell types 64
5.1.1 Prokaryotic hosts 64
5.1.2 Eukaryotic hosts 65
5.2 Plasmid vectors for use in E. coli 66
5.2.1 What are plasmids? 66
5.2.2 Basic cloning plasmids 67
5.2.3 Slightly more exotic plasmid vectors 69
5.3 Bacteriophage vectors for use in E. coli 70
5.3.1 What are bacteriophages? 71
5.3.2 Vectors based on bacteriophage 75
5.3.3 Vectors based on bacteriophage M13 78
5.4 Other vectors 79
5.4.1 Hybrid plasmid/phage vectors 80
5.4.2 Vectors for use in eukaryotic cells 80
5.4.3 Artificial chromosomes 83
5.5 Getting DNA into cells 84
5.5.1 Transformation and transfection 84
