An Introduction to Genetic Engineering

by Desmond S. T. Nicholl

Overview

The fourth edition of this popular textbook retains its focus on the fundamental principles of gene manipulation, providing an accessible and broad-based introduction to the subject for beginning undergraduate students. It has been brought thoroughly up to date with new chapters on the story of DNA and genome editing, and new sections on bioethics, significant developments in sequencing technology and structural, functional and comparative genomics and proteomics, and the impact of transgenic plants. In addition to chapter summaries, learning objectives, concept maps, glossary and key word lists the book now also features new concluding sections, further reading lists and web-search activities for each chapter to provide a comprehensive suite of learning resources to help students develop a flexible and critical approach to the study of genetic engineering.

Product Details

ISBN-13:	9781009190312
Publisher:	Cambridge University Press
Publication date:	03/02/2023
Sold by:	Barnes & Noble
Format:	eBook
File size:	39 MB
Note:	This product may take a few minutes to download.

About the Author

Desmond S. T. Nicholl was Senior Lecturer in Biological Sciences, Head of Bioscience, Head of Quality Enhancement and Assistant Dean for Education at the University of the West of Scotland. As well as three previous editions of An Introduction to Genetic Engineering, he also authored Cell and Molecular Biology (Learning & Teaching Scotland, 2000).

Table of Contents

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

Editorial Reviews

‘Genetic engineering represents a toolbox that all students within the basic and applied biology fields must get acquainted with. The fourth edition of An Introduction to Genetic Engineering is an excellent up-to-date version of a classic textbook. This ambitious book excellently balances the molecular biology knowledge required to grasp the comprehensive gene technology toolbox with a discussion of its impact on society.’ Per Amstrup Pedersen, University of Copenhagen

‘As a biomedical engineering professor teaching an undergraduate Genetic Engineering course for close to 10 years, I use Dr Nicholl’s An Introduction to Genetic Engineering as my go-to textbook. It is not one of those overly thick textbooks that overwhelm students. Its comprehensiveness captures readers’ attention with succinct fundamental concepts that truly promote one’s interest in exploring the wonder of many genetic engineering techniques and applications. To facilitate that further, the material provided at the end of each chapter encourages readers to expand their learning with relevant resources … Many of my students become so interested that they pursue graduate degrees and have a career in this field. Dr Nicholl’s textbook has a long-term influence on its readers.’ M. Ete Chan, State University of New York at Stony Brook

‘Dr Nicholl’s book covers all the basic material that one would expect from its title, but what particularly impressed me was how it isn’t afraid to move into political and socio-economic arenas. In Chapter 16, for example, balanced arguments are presented for and against the development of transgenic organisms, and these don’t always come out in favour of the science.’ Neil Crickmore, University of Sussex

From the Publisher