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.

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: J.A. Stader and T.J. Silhavy, Engineering Escherichia coli to Secrete Heterologous Gene Products. Protein Refolding: T. Kohno, D.F. Carmichael, A. Sommer, and R.C. Thompson, Refolding of Recombinant Proteins. Expression in Bacillus subtilis: D.J. Henner, Expression of Heterologous Genes in Bacillus subtilis. Intracellular Production: S.F.J. Le Grice, Regulated Promoter for High-Level Expression of Heterologous Genes in Bacillus subtilis. Secretion: V. Nagarajan, System for Secretion of Heterologous Proteins in Bacillus subtilis. Inducible Expression of Regulatory Proteins: D.J. Henner, Inducible Expression of Regulatory Genes in Bacillus subtilis. Expression in Yeast: S.D. Emr, Heterologous Gene Expression in Yeast. Plasmid Vectors: A.B. Rose and J.R. Broach, Propagation and Expression of Cloned Genes in Yeast: 2-µCircle-Based Vectors. T.Stearns, H. Ma, and D. Botstein, Manipulating Yeast Genome Using Plasmid Vectors. Inducible Expression Cassettes: L.M. Mylin, K.J. Hofmann, L.D. Schultz, and J.E. Hopper, Regulated GAL4 Expression Cassette Providing Controllable and High-Level Output from High-Copy Galactose Promoters in Yeast. V.L. Price, W.E. Taylor, W. Clevenger, M. Worthington, and E.T. Young, Expression of Heterologous Proteins in Saccharomyces cerevisiae Using ADH2 Promoter. T. Etcheverry, Induced Expression Using Yeast Copper Metallothionein Promoter. Constitutive Expression Cassettes: S.M. Kingsman, D. Cousens, C.A. Stanway, A. Chambers, M. Wilson, and A.J. Kingsman, High-Efficiency Yeast Expression Vectors Based on the Promoter of the Phosphoglycerate Kinase Gene. S. Rosenberg, D. Coit, and P. Tekamp-Olson, Glyceraldehyde-3-phosphate Dehydrogenase-Derived Expression Cassettes for Constitutive Synthesis of Heterologous Proteins. A.Z. Sledziewski, A. Bell, C. Yip, K. Kelsay, F.J. Grant, and V.L. MacKay, Superimposition of Temperature Regulation on Yeast Promoters. Translation: T.F. Donahue and A.M. Cigan, Sequence and Structural Requirements for Efficient Translation in Yeast. Proteases: E.W. Jones, Vacuolar Proteases in Yeast Saccharomyces cerevisiae. K.D. Wilkinson, Detection and Inhibition of Ubiquitin-Dependent Proteolysis. R.L. Kendall, R. Yamada, and R.A. Bradshaw, Cotranslational Amino-Terminal Processing. Protein Secretion and Modification: A.J. Brake, à-Factor Leader-Directed Secretion of Heterologous Proteins from Yeast. R.A. Hitzeman, C.Y. Chen, D.J. Dowbenko, M.E. Renz, C. Liu, R. Pai, N.J. Simpson, W.J. Kohr, A. Singh, V. Chisholm, R. Hamilton, and C.N. Chang, Use of Heterologous and Homologous Signal. Sequences for Secretion of Heterologous Proteins from Yeast. C.E. Ballou, Isolation, Characterization, and Properties of Saccharomyces cerevisiae mnn Mutants with Nonconditional Protein Glycosylation Defects. V. Chisholm, C.Y. Chen, N.J. Simpson, and R.A. Hitzeman, Molecular and Genetic Approach to Enhancing Protein Secretion. Expression in Mammalian Cells: A.D. Levinson, Expression of Heterologous Genes in Mammalian Cells. Vectors: R.J. Kaufman, Vectors Used for Expression in Mammalian Cells. M. Kriegler, Assembly of Enhancers, Promoters, and Splice Signals to Control Expression of Transferred Genes. Transfection Methods: W.A. Keown, C.R. Campbell, and R.S. Kucherlapati, Methods for Introducing DNA into Mammalian Cells. Markers for Selection and Amplification: R.J. Kaufman, Selection and Coamplification of Heterologous Genes in Mammalian Cells. Growth of Cell Lines: J.P. Mather, Optimizing Cell and Culture Environment for Production of Recombinant Proteins. Posttranslation Processing, Modification, and Secretion: A.J. Dorner and R.J. Kaufman, Analysis of Synthesis, Processing, and Secretion of Proteins Expressed in Mammalian Cells. Mutagenesis: B.H. Horwitz and D. DiMaio, Saturation Mutagenesis Using Mixed Oligonucleotides and M13 Templates Containing Uracil. Author Index. Subject Index.