The methods presented in this volume will enable the reader to design effective strategies for the expression of cloned genes and cDNAs and will prove useful in solving the majority of expression problems one is likely to encounter.


Biochemists, geneticists, microbiologists, genetic engineers, analytical chemists, molecular biologists, biotechnologists, and industrial researchers working on protein products.

Table of Contents

Introduction: D.V. Goeddel, Systems for Heterologous Gene Expression. Expression in Escherichia coli: L. Gold, Expression of Heterologous Proteins in Escherichia coli. Vectors: P. Balbas and F. Bolivar, Design and Construction of Expression Plasmid Vectors in Escherichia coli. Promoters: J.D. Gralla, Promoter Recognition and mRNA Initiation by Escherichia coli Eo-70. D.G. Yansura and D.J. Henner, Use of Escherichia coli trp Promoter for Direct Expression of Proteins. F.W. Studier, A.H. Rosenberg, J.J. Dunn, and J.W. Dubendorff, Use of T7 RNA Polymerase to Direct Expression of Cloned Genes. Translation: L. Gold and G.D. Stormo, High-Level Translation Initiation. B.E. Schoner, R.M. Belagaje, and R.G. Schoner, Enhanced Translational Efficiency with Two-Cistron Expression System. H.A. de Boer and A.S. Hui, Sequences within Ribosome Binding Site Affecting Messenger RNA Translatability and Method to Direct Ribosomes to Single Messenger RNA Species. P.O. Olins and S.H. Rangwala, Vector for Enhanced Translation of Foreign Genes in Escherichia coli. Proteases: S. Gottesman, Minimizing Proteolysis in Escherichia coli: Genetic Solutions. Gene Fusions: M. Uhlén and T. Moks, Gene Fusions for Purpose of Expression: An Introduction. B. Nilsson and L. Abrahmsén, Fusions to Staphylococcal Protein A. D.G. Yansura, Expression as trpE Fusion. Secretion:


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© 1990
Academic Press
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@qu:"The purpose of this volume is to give recombinant DNA researchers the most up-to-date knowledge about heterologous gene expression in four commonly employed organisms: Escherichia coli, Bacillus subtilis, yeast, and mammalian cells....This synthesis of theory and practice has, in general, been skillfully done, making this volume most worthwhile for students and researchers in this area. Expert coverage in each of the four organisms ensures that the knowledge and advice given in each area is first rate....This volume should prove invaluable to investigators with a molecular biology background who need to express a given gene, but are not particularly experienced in this area. Many of the contributors have been able to effectively communicate years worth of experience and unpublished observations in what often was a black magic science. This volume clearly shows that things have changed and that the methodology is now in the hands of the skilled non-expert." @source:--AMERICAN JOURNAL OF PHYSIOLOGY @qu:"The volume is very successful...I strongly recommend this book both to novices who are embarking on a gene expression project for the first time and to frustrated veterans who are seeking the latest 'tips' about how to improve the expression of their favourite proteins." @source:--TRENDS IN GENETICS @qu:"This volume presents an extremely valuable collection of methods for expression of cloned genes...This volume fills an important void in the scientific literature. It is designed to be an integrated treatment of expression systems rather than a collection of relevant papers. In most respects it accomplishes that goal admirably and it provides an invaluable resource that should be present in any laboratory interested in manipulating gene expression." @source:--ANALYTICAL BIOCHEMISTRY @qu:"This work is essential for the experienced worker or novice who having cloned his gene wishes to expr