Bibliographic record and links to related information available from the Library of Congress catalog.
Note: Contents data are machine generated based on pre-publication provided by the publisher. Contents may have variations from the printed book or be incomplete or contain other coding.
Contents Chapter 1: Introduction 1.1 The beginning of gene cloning 1.2 How to use this book 1.3 What you need to know before you read this book 1.4 A request from the authors Further reading Chapter 2: Genome organization 2.1 Introduction 2.2 The C-value paradox 2.3 The human genome 2.4 Genomes of other eukaryotes 2.5 Bacterial genomes 2.6 Plasmids 2.7 Viral genomes 2.8 GC content 2.9 Physical characteristics of eukaryotic chromosomes 2.10 Karyotype 2.11 Euchromatin and heterochromatin 2.12 CpG islands Questions and answers Further reading Chapter 3: Key tools for gene cloning 3.1 Introduction 3.2 Vectors 3.3 Restriction enzymes 3.4 DNA ligase 3.5 Transformation 3.6 Purification of plasmid DNA 3.7 More restriction enzymes 3.8 Alkaline phosphatase 3.9 More about vectors 3.10 Analysing cloned DNA by restriction mapping 3.11 Measuring the size of DNA fragments 3.12 The polymerase chain reaction and its use in gene cloning 3.13 How does PCR work? 3.14 Designing PCR primers 3.15 The PCR reaction 3.16 Uses for PCR products 3.17 Cloning PCR products 3.18 Real-time PCR for quantification of DNA 3.19 Advantages and limitations of PCR Questions and answers Further reading Chapter 4: Gene identification and DNA libraries 4.1 The problem 4.2 Genomic library 4.3 Constructing a genomic library 4.4 How many clones? 4.5 Some DNA fragments are under-represented in genomic libraries 4.6 Using partial digests to make a genomic library 4.7 Storage of genomic libraries 4.8 Advantages and disadvantages of genomic libraries 4.9 Cloning vectors for gene libraries 4.10 Vectors derived from bacteriophage 4.11 Packing bacteriophage in vitro 4.12 Cloning with bacteriophage 4.13 Calculating the titer of your library 4.14 Cosmid libraries 4.15 Making a cosmid library 4.16 YAC and BAC vectors 4.17 cDNA libraries 4.18 Making a cDNA library 4.19 Cloning the cDNA product 4.20 Expressed sequence tags 4.21 What are the disadvantages of a cDNA library? Questions and answers Further reading Chapter 5: Screening DNA libraries 5.1 The problem 5.2 Screening methods based on gene expression 5.3 Complementation 5.4 Immunological screening of expression libraries 5.5 Screening methods based on detecting a DNA sequence 5.6 Oligonucleotide probes 5.7 Cloned DNA fragments as probes 5.8 Colony and plaque hybridization 5.9 Differential screening 5.10 Using PCR to screen a library Questions and answers Further reading Chapter 6: Further routes to gene identification 6.1 How do we get from phenotype to gene: a fundamental problem in gene cloning 6.2 Gene tagging: a method that both mutates and marks genes 6.3 A simple example of transposon tagging in bacteria: cloning adhesive genes from pseudomonas 6.4 Signature-tagged mutagenesis: cloning bacterial genes with "difficult" phenotypes 6.5 Gene tagging in higher eukaryotes: resistance genes in plants 6.6 Positional cloning: using maps to track down genes 6.7 Identification of a linked marker 6.8 Moving from the marker towards the gene of interest 6.9 Identifying the gene of interest 6.10 Cloning of the CF gene: a case study Questions and answers Further reading Chapter 7: Sequencing DNA 7.1 Introduction 7.2 Overview of sequencing 7.3 Sanger sequencing 7.4 The Sanger sequencing protocol requires a single-stranded DNA template 7.5 Modifications of the original Sanger protocol 7.6 Strategies for sequencing a DNA fragment 7.7 High-throughput sequencing protocols 7.8 The modern sequencing protocol 7.9 Genome sequencing 7.10 High-throughput pyrosequencing 7.11 The importance of DNA sequencing Questions and answers Further reading Chapter 8: Bioinformatics 8.1 Introduction 8.2 What does a gene look like? 8.3 Identifying eukaryotic genes 8.4 Sequence comparisons 8.5 Pair-wise comparisons 8.6 Identity and similarity 8.7 Is the alignment significant? 8.8 What can alignments tell us about the biology of the sequences being compared? 8.9 Similarity searches 8.10 Fasta 8.11 BLAST 8.12 What can similarity searches tell us about the biology of the sequences being compared? 8.13 Multiple sequence alignments 8.14 What can multiple sequence alignments tell us about the structure and function of proteins? 8.15 Consensus patterns and sequence motifs 8.16 Investigating the three-dimensional structures of biological molecules 8.17 Using sequence alignments to create a phylogenetic tree Questions and answers Further reading Chapter 9: Production of proteins from cloned genes 9.1 Why express proteins? 9.2 Requirements for protein production from cloned genes 9.3 The use of E. coli as a host organism for protein production 9.4 Some problems in obtaining high level production of proteins in E. coli 9.5 Beyond E. coli: protein expression in eukaryotic systems 9.6 A final word about protein purification Questions and answers Further reading Chapter 10: Gene cloning in the functional analysis of proteins 10.1 Introduction 10.2 Analyzing the expression and role of unknown genes 10.3 Determining the cellular location of proteins 10.4 Mapping of membrane proteins 10.5 Detecting interacting proteins 10.6 Site-directed mutagenesis for detailed probing of gene and protein function Questions and answers Further reading Chapter 11: The analysis of the regulation of gene expression 11.1 Introduction 11.2 Determining the transcription start of a gene 11.3 Determining the level of gene expression 11.4 Identifying the important regulatory regions 11.5 Identifying protein factors 11.6 Global studies of gene expression Questions and answers Further reading Chapter 12: The production and uses of transgenic organisms 12.1 What is a transgenic organism? 12.2 Why make transgenic organisms? 12.3 How are transgenic organisms made? 12.4 Drawbacks and problems 12.5 Knockout mice and other organisms: the growth of precision in transgene targeting 12.6 Is the technology available to produce transgenic people? Questions and answers Further reading Chapter 13: Forensic and medical applications 13.1 Introduction 13.2 Forensics 13.3 DNA profiling 13.4 Multiplex PCR 13.5 Samples for forensic analysis 13.6 Obtaining more information from DNA profiles 13.7 Other applications of DNA profiling 13.8 Medical applications 13.9 Techniques for diagnosis of inherited disorders 13.10 Whole genome amplification 13.11 Diagnosis of infectious disease 13.12 Diagnosis and management of cancer Questions and answers Further reading
Library of Congress Subject Headings for this publication:
Molecular cloning.
Cloning, Molecular.
Gene Library.
Genomics -- methods.