Brown - Gene cloning and DNA analysis: an introduction

T.A. BROWN

University of Manchester
Manchester

Seventh Edition

This edition first published 2016 2016 by John Wiley & Sons, Ltd
Sixth edition published 2010 by John Wiley & Sons, Ltd
Fourth and fifth editions published 2001, 2006 by Blackwell Publishing Ltd
First, second and third editions published 1986, 1990, 1995 by Chapman & Hall

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Library of Congress Cataloging-in-Publication Data

Brown, T.A. (Terence A.), author.
Gene cloning and DNA analysis : an introduction / T.A. Brown. Seventh edition.
p. ; cm.
Includes bibliographical references and index.
ISBN 978-1-119-07257-7 (cloth) ISBN 978-1-119-07256-0 (pbk.)
I. Title.
[DNLM: 1. Cloning, Molecular. 2. DNA, Recombinantanalysis. 3. Sequence Analysis, DNA. QU 450]
QH442.2
572.8633dc23

2015015462

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Anyone who works with DNA is well aware of the dramatic changes that have taken place during the past few years in DNA sequencing methodology. To reflect these advances, in this new edition of Gene Cloning and DNA Analysis: An Introduction I have completely remodelled the chapter on DNA sequencing to give the new next-generation methods equal prominence alongside the traditional approaches to DNA sequencing, and also to modernize the description of the ways in which genome sequences are generated. Elsewhere, I have stressed the importance of RNA-seq as a means of studying transcriptomes, and ChIP-seq for locating protein-binding sites. These changes correct the major weakness of the Sixth Edition, which was written just before these methods came into mainstream use.

Elsewhere, I have made the usual updates, especially in Part III where I have tried to keep pace with the increasingly rapid developments in the applications of gene cloning and DNA analysis in industry, medicine and agriculture. I have also rewritten the last part of the final chapter, on archaeogenetics, in order to present some of the new information on the human past that has been revealed by the Neanderthal and Denisovan genome sequences. As always, my primary aim is to ensure that Gene Cloning remains an introductory text that begins at the beginning and does not assume that the reader has any prior knowledge of the techniques used to study genes and genomes.

For the n-th time I must thank my wife Keri for the unending support that she has given to me in my decision to use up evenings and weekends writing this and other books.

T.A. Brown

University of Manchester

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In the middle of the 19th century, Gregor Mendel formulated a set of rules to explain the inheritance of biological characteristics. The basic assumption of these rules is that each heritable property of an organism is controlled by a factor, called a gene , that is a physical particle present somewhere in the cell. The rediscovery of Mendel's laws in 1900 marks the birth of genetics , the science aimed at understanding what these genes are and exactly how they work.

For the first 30 years of its life this new science grew at an astonishing rate. The idea that genes reside on chromosomes was proposed by W. Sutton in 1903, and received experimental backing from T.H. Morgan in 1910. Morgan and his colleagues then developed the techniques for gene mapping , and by 1922 had produced a comprehensive analysis of the relative positions of over 2000 genes on the four chromosomes of the fruit fly, Drosophila melanogaster.

Despite the brilliance of these classical genetic studies, there was no real understanding of the molecular nature of the gene until the 1940s. Indeed, it was not until the experiments of Avery, MacLeod, and McCarty in 1944, and of Hershey and Chase in 1952, that anyone believed that deoxyribonucleic acid (DNA) is the genetic material. Up until then it was widely thought that genes were made of protein. The discovery of the role of DNA was a tremendous stimulus to genetic research, and many famous biologists (Delbrck, Chargaff, Crick, and Monod were among the most influential) contributed to the second great age of genetics. During the 14 years between 1952 and 1966, the structure of DNA was elucidated, the genetic code cracked, and the processes of transcription and translation described.

These years of activity and discovery were followed by a lull, a period of anticlimax when it seemed to some molecular biologists (as the new generation of geneticists styled themselves) that there was little of fundamental importance that was not understood. In truth, there was a frustration that the experimental techniques of the late 1960s were not sophisticated enough to allow the gene to be studied in any greater detail.