Genetic Engineering with PCR Book Reviews
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Review by April 1998
This multi-author book describes genetic engineering techniques using the polymerase chain reaction (PCR) in the following chapters: (1) introduction to genetic engineering with PCR; (2) mutagenesis by megaprimer PCR; (3) effects of dNTP and divalent metal ion concentration on random PCR mutagenesis; (4) rapid cycle amplification for construction of competitive templates; (5) DNA splicing by directed ligation; (6) multimers of specific DNA sequences generated by PCR; (7) use of PCR-based scanning mutagenesis in molecular cell biology; (8) in vitro synthesis of viroids; (9) in vitro selection of functional nucleic acid sequences; (10) directed and random recombination of antibody sequences; (11) construction, assembly and selection of combinatorial antibody libraries; and (12) beyond the limits of natural diversity: PCR synthesis of semi-random peptide and antibody phage display libraries. An appendix outlining the basic technique of PCR is included as well as a list of standard abbreviations for ambiguous nucleic acid sequences.
Review by Genetical Research, August 1998 This book is a very useful addition to the growing number of 'textbooks' dealing with the polymerase chain reaction (PCR). It has several features that distinguish it from 'run of the mill' PCR books. The first chapter (which is written by the editors) is a well considered and thought provoking discourse on the position of PCR in the development of genetic engineering. The authors recognize three generations of DNA technology: 1) conventional DNA genetics; 2) recombinant DNA cloning; 3) PCR-based DNA manipulation in vitro. They argue that PCR will be as important to the development of gene manipulation as the first two approaches. The rest of the book deals with PCR methodology, but it is not a 'cookbook' although some of the chapters do provide fairly detailed protocols. Each chapter has, in any case, such a comprehensive list of references that the reader is easily able to obtain all of the essential experimental details. The special merits of this text include the clear layout, the subject areas covered and the selection of such appropriate examples to illustrate techniques as to ensure wide applicability. Although this is a multi-authored work composed of chapters which cover divergent aspects of PCR, the text is so effectively constructed and edited that it is well integrated. The editors have resisted the temptation to have several chapters dealing with the basics of PCR (an appendix gives some background information which, considering the complexity of some of the material in the book, is no bad thing). Some pearls of wisdom that are applicable to all aspects of PCR are to be found in the book. Following the opening chapter there are, for instance, chapters dealing with the fidelity of PCR amplification and the effects of dNTP concentration and divalent ion composition, rapid cycle amplification, mutagenesis and DNA splicing by SDL, which will find wide applicability amongst genetic engineers. These chapters also provide a useful background for some of those that follow. Later chapters cover topics such as scanning mutagenesis (homology scanning and alanine scanning) in order to identify important amino acids or regions in proteins. This chapter also deals with methods for inserting 'tags' or 'handles' into non-crucial areas of proteins to aid identification or purification. Several chapters cover the exciting areas of molecular evolution and/or antibody engineering from a variety of conceptual and methodological backgrounds (SELEX, phage display, etc.). These chapters are fairly complex and specialized. This book is commendably up-to-date and covers areas not usually provided in books of this type and price. It certainly justifies its inclusion in the Current Innovations in Molecular Biology series. The book is not for beginners but we strongly recommend it to scientists who anticipate sophisticated exploitation of PCR in genetic engineering.
Review by Human Genetics 102(6); 706, July 1998 The first good thing about this book is that it is quite obvious, right from the start, that it is not an introductory text for PCR beginners. It is therefore, quite sensible not to bother an "advanced" reader with an introductory chapter about the basic principles of PCR, but rather to put this chapter right at the end of the book. There it is camouflaged as an Appendix quite appropriately named "A primer on PCR". The benefits of this elegant structural solution, however are partially undermined by the inclusion of both a preface, and the first chapter "An introduction to genetic engineering with PCR". This, I'm afraid, is not the only problem with the organization of the text. If you were to read this book from A to Z (as only reviewers are supposed to), you would find a lot of theory and previews to the interesting chapters in the middle of the book. Unfortunately, you would not really know what "sexual PCR" was before you read the corresponding chapter. It is quite clear from the outset that this is not a book on analytical PCR with all the diagnostic and sensitivity problems amply described elsewhere. It might have been a very good reference text for those with a specific "engineering" or "construction" PCR problem, except that its format does not offer ready guidance to those seeking solutions. This might well be the weakest point of the book, especially since some of the chapter headings disguise their actual content. For instance, hidden in Chapter 4 ("Rapid cycle amplification for construction of competitive templates") you will find a very useful recipe for the construction of insertion products, which would hardly been expected from the chapter's title. Unfortunately the Index does not offer any assistance here, since it doesn't have an entry for "insertions" either. If you make your way to the central chapters of the book, you will be taken by surprise at just how far the PCR technology has developed, even if you are a PCR professional. Here you will read about mutagenesis, multimerization, in vitro synthesis and selection of nucleic acids. Another highlight awaits the reader in Chapter 7 ("The use of PCR-based scanning mutagenesis in molecular cell biology"), but again, the chapter title successfully hides its content. Here the possibility of functional sequence is described in a way molecular geneticists might only have dreamed of ten years ago.
In summary, this book contains plenty of useful technological information, but you have to be prepared to search for it carefully. At the end of the book, you are left wondering why we need not one, nor two, but three chapters on the construction of antibody libraries. At the end of this review you can be assured that this is the right book for all those who think they know everything there is to know about PCR- or have such a student in their seminars.
Review by SGM Quarterly/Microbiology Today 25(4); 170, November 1998 For those who consider PCR only to be an analytical tool then think again.This book explores technical aspects of DNA manipulation using PCR rather than traditional cloning methods.Topics include mutagenesis (targeted and random), recombination, splicing, selection of functional nucleic acids, antibody and phage display libraries and viroid synthesis. Additionally, a chapter about controlling Taq -induced errors is essential reading for those wanting to reduce or maintain the sequence fidelity of the final product.The concept of rapid PCR (30 cycles in 15 minutes) using hot-air cyclers also features. PCR-meditated genetic engineering can have advantages over traditional cloning methods.The whole process can be carried out in vitro without need to amplify constructs in vivo . Consequently there are significant time savings and an ability to generate constructs that would otherwise be toxic, unstable or unclonable previously. An excellent read for those familiar with traditional molecular biology. Current Microbiology 38: 312. August 1999 This paperback book is the fifth and most recent volume in the series Current Innovations in Molecular Biology. The hardback copy is available at $109.99. This well-done book is not so much a how-to manual as one that provides the reader with more than adequate "how it is done" and readily understood information from the real world of practical science. An assemblage of molecular scientists from around the globe authored the 12 chapters and covered in detail a large variety of functional aspects of genetic modification or engineering. The various chapters address mutagenesis by megaprimer PCR, effect of divalent metal ion concentration on PCR mutagenesis, rapid cycle amplification for competitive templates, synthesis of viroids, and looking beyond the limits of natural diversity. There are many other equally important issues addressed; the above listing provides only a sample. Is it easy to understand and comprehend? It is well written, but in my opinion easily understood only by the well-informed molecular biologist or genetic manipulator. Understanding the content of this book is eased to some extent by an appendix titled "A Primer on PCR," but to many it is a format more readily digested by the fully informed person who desires to polish her or his skills and knowledge. As mentioned above, this is not a how to book, but is one that will be referred to often and should be readily at hand for the molecular biologist seeking new avenues and manipulations to pursue in genetic engineering using PCR.
CAB International
Mark R. Fowler, Nigel W. Scott, Malcolm C. Elliott The Norman Borlaug Institute for Plant Science Research, De Montfort University, Scraptoft, Leicester, LE7 9SU, U.K.
Jochen Reiss Institute for Human Genetics, Gottingen, Germany.
Barry Vipond Public Health Laboratory, Bristol, UK
Review by
W. J. Hausler Jr. Iowa City, IA, USA