From Genes to Genomes, 3ed: Concepts and Applications of DNA Technology

Jeremy W. Dale, Malcolm von Schantz, Nicholas Plant

ISBN: 9788126598083

400 pages

Exclusively distributed by CBS Publishers & Distributors

 

Description

Rapid advances in a collection of techniques referred to as gene technology, genetic engineering, recombinant DNA technology and gene cloning have pushed molecular biology to the forefront of the biological sciences. From Genes to Genomes: Concepts and Applications of DNA Technology explains key ideas underlying the most central techniques in the context of the ways in which they are used. The book opens with a brief review of the basic concepts of molecular biology, before moving on to describe the key molecular methods and how they fit together. This ranges from the cloning and study of individual genes to the sequencing of whole genomes, and the analysis of genome-wide information.

Preface

 

1 From Genes to Genomes

1.1 Introduction  

1.2 Basic molecular biology  

1.3 What is a gene?

1.4 Information flow: gene expression

1.5 Gene structure and organisation

1.6 Refinements of the model

 

2 How to Clone a Gene

2.1 What is cloning?

2.2 Overview of the procedures

2.3 Extraction and purification of nucleic acids

2.4 Detection and quantitation of nucleic acids

2.5 Gel electrophoresis

2.6 Restriction endonucleases

2.7 Ligation

2.8 Modification of restriction fragment ends

2.9 Plasmid vectors

2.10 Vectors based on the lambda bacteriophage

2.11 Cosmids

2.12 Supervectors: YACs and BACs

2.13 Summary

 

3 Genomic and cDNA Libraries

3.1 Genomic libraries

3.2 Growing and storing libraries

3.3 cDNA libraries

3.4 Screening libraries with gene probes

3.5 Screening expression libraries with antibodies

3.6 Characterization of plasmid clones

 

4 Polymerase Chain Reaction (PCR)

4.1 The PCR reaction

4.2 PCR in practice

4.3 Cloning PCR products

4.4 Long-range PCR

4.5 Reverse-transcription PCR

4.6 Quantitative and real-time PCR

4.7 Applications of PCR

 

5 Sequencing a Cloned Gene

5.1 DNA sequencing

5.2 Databank entries and annotation

5.3 Sequence analysis

5.4 Sequence comparisons

5.5 Protein structure

5.6 Confirming gene function

 

6 Analysis of Gene Expression

6.1 Analysing transcription

6.2 Methods for studying the promoter

6.3 Regulatory elements and DNA-binding proteins

6.4 Translational analysis

 

7 Products from Native and Manipulated Cloned Genes

7.1 Factors affecting expression of cloned genes

7.2 Expression of cloned genes in bacteria

7.3 Yeast systems

7.4 Expression in insect cells: baculovirus systems

7.5 Mammalian cells

7.6 Adding tags and signals

7.7 In vitro mutagenesis

7.8 Vaccines

 

8 Genomic Analysis

8.1 Overview of genome sequencing

8.2 Next generation sequencing (NGS)

8.3 De novo sequence assembly

8.4 Analysis and annotation

8.5 Comparing genomes

8.6 Genome browsers

8.7 Relating genes and functions: genetic and physical maps

8.8 Transposon mutagenesis and other screening techniques

8.9 Gene knockouts, gene knockdowns and gene silencing

8.10 Metagenomics

8.11 Conclusion

 

9 Analysis of Genetic Variation

9.1 Single nucleotide polymorphisms

9.2 Larger scale variations

9.3 Other methods for studying variation

9.4 Human genetic variation: relating phenotype to genotype

9.5 Molecular phylogeny

 

10 Post-Genomic Analysis

10.1 Analysing transcription: transcriptomes

10.2 Array-based methods

10.3 Transcriptome sequencing

10.4 Translational analysis: proteomics

10.5 Post-translational analysis: protein interactions

10.6 Epigenetics

10.7 Integrative studies: systems biology

 

11 Modifying Organisms: Transgenics

11.1 Transgenesis and cloning

11.2 Animal transgenesis

11.3 Applications of transgenic animals

11.4 Disease prevention and treatment

11.5 Transgenic plants and their applications

11.6 Transgenics: a coda

 

Glossary

Bibliography

Index