Gene Cloning and DNA Analysis addresses new and growing areas of research whilst retaining the philosophy of the previous editions. Assuming the reader has little prior knowledge of the subject, its importance, the principles of the techniques used and their applications are all carefully laid out, with over 250 clearly presented four-colour illustrations.

Contents

Part 1: The Basic Principles of Gene Cloning and DNA Analysis

• Why Gene Cloning and DNA Analysis are Important
• The early development of genetics
• The advent of gene cloning and the polymerase chain reaction
• What is gene cloning
• What is PCR
• Why gene cloning and PCR are so important
• How to find your way through this book

Part 2: Vectors for Gene Cloning: Plasmids and Bacteriophages

• Plasmids
• Bacteriophages

Part 3: Purification of DNA from Living Cells

• Preparation of total cell DNA
• Preparation of plasmid DNA
• Preparation of bacteriophage DNA

Part 4: Manipulation of Purified DNA

• The range of DNA manipulative enzymes
• Enzymes for cutting DNA – restriction endonucleases
• Ligation – joining DNA molecules together

Part 5: Introduction of DNA into Living Cells

• Transformation – the uptake of DNA by bacterial cells
• Identification of recombinants
• Introduction of phage DNA into bacterial cells
• Identification of recombinant phages
• Introduction of DNA into non-bacterial cells

Part 6: Cloning Vectors for E. coli

• Cloning vectors based on E. coli plasmids
• Cloning vectors based on M13 bacteriophage
• Cloning vectors based on l bacteriophage
• λ and other high capacity vectors enable genomic libraries to be constructed
• Vectors for other bacteria

Part 7: Cloning Vectors for Eukaryotes

• Vectors for yeast and other fungi
• Cloning vectors for higher plants
• Cloning vectors for animals

Part 8: How to Obtain a Clone of a Specific Gene

• The problem of selection
• Direct selection
• Identification of a clone from a gene library
• Methods for clone identification

Part 9: The Polymerase Chain Reaction

• The polymerase chain reaction in outline
• PCR in more detail
• After the PCR: studying PCR products
• Real time PCR enables the amount of starting material to be quantified

Part 10: Sequencing Genes and Genomes

• The methodology for DNA sequencing
• How to sequence a genome

Part 11: Studying Gene Expression and Function

• Studying the RNA transcript of a gene
• Studying the regulation of gene expression
• Identifying and studying the translation product of a cloned gene

Part 12: Studying Genomes

• Genome annotation
• Studies of the transcriptome and proteome

Part 13: Production of Protein from Cloned Genes

• Special vectors for expression of foreign genes in E. coli
• General problems with the production of recombinant protein in E. coli
• Production of recombinant protein by eukaryotic cells

Part 14: Gene Cloning and DNA Analysis in Medicine

• Production of recombinant pharmaceuticals
• Identification of genes responsible for human diseases
• Gene therapy

Part 15: Gene Cloning and DNA Analysis in Agriculture

• The gene addition approach to plant genetic engineering
• Gene subtraction
• Problems with genetically modified plants

Part 16: Gene Cloning and DNA Analysis in Forensic Science and Archaeology

• DNA analysis in the identification of crime suspects
• Studying kinship by DNA profiling
• Sex identification by DNA analysis
• Archaeogenetics – using DNA to study human prehistory

Glossary

Index

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