前辅文
Part I The Basic Principles of Gene Cloning and DNA Analysis
1 Why Gene Cloning and DNA Analysis are Important
1.1 The early development of genetics
1.2 The advent of gene cloning and the polymerase chain reaction
1.3 What is gene cloning?
1 .4 What is PCR?
1.5 Why gene cloning and PCR are so important
1.6 How to find your way through this book
2 Vectors for Gene Cloning: Plasmids and Bacteriophages
2.1 Plasmids
2.2 Bacteriophages
2.2.3 Viruses as cloning vectors for other organisms
3 Purification of DNA from Living Cells
3.1 Preparation of total cell DNA
3.2 Preparation of plasmid DNA
3.3 Preparation of bacteriophage DNA
4 Manipulation of Purified DNA
4.1 The range of DNA manipulative enzymes
4.2 Enzymes for cutting DNA: Restriction endonucleases
4.3 Ligation: Joining DNA molecules together
5 Introduction of DNA into Living Cells
5.1 Transformation: The uptake of DNA by bacterial cells
5.2 Identification of recombinants
5.3 Introduction of phage DNA into bacterial cells
5.4 Identification of recombinant phages
5.5 Introduction of DNA into non-bacterial cells
6 Cloning Vectors for Escherichia co/i
6.1 Cloning vectors based on E. co/i plasmids
6.2 Cloning vectors based on à bacteriophage
6.3 Cloning vectors for the synthesis of single-stranded DNA
6.4 Vectors for other bacteria
7 Cloning Vectors for Eukaryotes
7.1 Vectors for yeast and other fungi
7.2 Cloning vectors for higher plants
7.3 Cloning vectors for animals
8 How to Obtain a Clone of a Specific Gene
8.1 The problem of selection
8.2 Direct selection
8.3 Identification of a clone from a gene library
8.4 Methods for clone identification
9 The Polymerase Chain Reaction
9.1 PCR in outline
9.2 PCR in more detail
9.3 After the PCR: Studying PCR products
9.4 Real-time PCR enables the amount of starting material to be quantified
Part 11 The Applications of Gene Cloning and DNA Analysis in Research
10 Sequencing Genes and Genomes
10.1 Chain-termination DNA sequencing
10.2 Next-generation sequencing
10.3 How to sequence a genome
11 Studying Gene Expression and Function
11.1 Studying the RNA transcript of a gene
11.2 Studying the regulation of gene expression
11.3 Identifying and studying the translation product of a cloned gene
12 Studying Genomes
12.1 Genome annotation
12.2 Studies of the transcriptome and proteome
Part 111 The Applications of Gene Cloning and DNA Analysis in Biotechnology
13 Production of Protein from Cloned Genes
13.1 Special vectors for the expression of foreign genes in E. co/i
13.2 General problems with the production of recombinant protein in E. co/i
13.3 Production of recombinant protein by eukaryotic cells
14 Gene Cloning and DNA Analysis in Medicine
14.1 Production of recombinant pharmaceuticals
14.2 Identification of genes responsible for human diseases
15 Gene Cloning and DNA Analysis in Agriculture
15.1 The gene addition approach to plant genetic engineering
15 . 2 Gene subtraction
15.3 Problems with genetically modified plants
16 Gene Cloning and DNA Analysis in Forensic Science and Archaeology
16.1 DNA analysis in the identification of crime suspects
16.2 Studying kinship by DNA profiling
16.3 Sex identification by DNA analysis
16.4 Archaeogenetics: Using DNA to study human prehistory
Glossary
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