Recombinant Gene Expression Protocols

Part I. Introduction. Recombinant Gene Expression Protocols: Overview of Experimental Strategies for the Expression of Recombinant Genes, Rocky S. Tuan. Part II. Expression in E. coli. Designing Expression Plasmid Vectors in E. coli, Paulina Balbas. Design of Bacterial Hosts for lac-Based Expression Vectors, Herbert P. Schweizer and RoxAnn R. Karkhoff-Schweizer. Bacteriophage l-Based Expression Vectors, Alan C. Christensen. Strategies for Gene Fusions, Stefan Stahl, Per-Ake Nygren, and Mathias Uhlen. Application of the E. coli trp Promoter, Daniel G. Yansura and Steven H. Bass. Expression of Proteins in E. coli Utilizing a Dual Promoter-Based Vector: pLACT7, George A. Garcia and Shaorong Chong. Expression and Secretion of Proteins in E. coli, Ophry Pines and Masayori Inouye. Synthetic Two-Cistron Expression System, Brigitte E. Schoner. Expression of cDNA-Encoded Proteins by Cell-Free Transcription and Translation, Christopher Coleclough. Part III. Expression in Yeast. Yeast Plasmids, Keshav K. Singh and Jack A. Heinemann. Inducible Expression Cassettes in Yeast: GAL4, Lawrence M. Mylin and James E. Hopper. Inducible Expression Cassettes in Yeast: ADH2, Virginia L. Price. Constitutive Expression Vectors: PGK, Ian R. Graham and Alistair Chambers. Part IV. Viral Expression Systems. Design and Construction of Recombinant Vaccinia Viruses, Christopher C. Broder and Patricia L. Earl. A Simplified Method of Screening for Isolation of Recombinant Vaccinia Virus, Haifeng Chen and R. Padmanabhan. Design of Retroviral Expression Vectors, Kenine E. Comstock, Nathaniel F. Watson, and John C. Olsen. Use of Defective Herpes-Derived Plasmid Vectors, Filip Lim, Dean Hartley, Philip Starr, Song Song, Phung Lang, Linda Yu, Yaming Wang, and Alfred I. Geller. Part V. Expression in Nonmammalian Higher Eukaryotes. Use of Baculovirus Expression Vectors, David R. O'Reilly. Expression of Exogenous Genes in Xenopus Oocytes, Eggs, and Embryos, Koichiro Shiokawa, Chie Koga, Yuzuru Ito, andMikihito Shibata. Analysis of Heterologous Gene Expression in Xenopus Blastomeres, Sally A. Moody. Part VI. Expression in Mammalian Cells. Overview of Vector Design for Mammalian Gene Expression, Randal J. Kaufman. Experimental Strategies in Efficient Transfection of Mammalian Cells: Calcium Phosphate and DEAE-Dextran, Gregory S. Pari and Wayne A. Keown. Experimental Strategies in Efficient Transfection of Mammalian Cells: Electroporation, Donald C. Chang. Highly Effective Delivery of Foreign DNA to Adherent Cells via Polybrene/DMSO-Assisted Gene Transfer, Remy A. Aubin, Michael Weinfeld, Marzieh Taghavi, Razmik Mirzayans, and Malcolm C. Paterson. Selection of Transfected Cells: Magnetic Affinity Cell Sorting, Raji Padmanabhan, Snorri S. Thorgeirsson, and R. Padmanabhan. Selection of Transfected Cells and Coamplification of Transfected Genes, Susan E. Kane. Optimization of Growth, Viability, and Specific Productivity for Expression of Recombinant Proteins in Mammalian Cells, Jennie P. Mather, Alison Moore, and Robert Shawley. Part VII. Expression in Transgenic Organisms. Promoters to Express Cloned Genes Uniformly in Drosophila, Jean-Paul Vincent and Charles Girdham. Application of Micromechanical Piercing Structures for Genetic Transformation of Nematodes, Sarwar Hashmi and Randy Gaugler. Strategies in Generating Transgenic Mammals, Olena Jacenko. Use of Gonadal Primordial Germ Cells (PGCs) as Tools for Gene Transfer in Chickens, Nathalie Allioli, Gerard Verdier, and Catherine Legras. Strategies for the Production of Transgenic Chickens, Robert J. Etches and Ann M. Verrinder Gibbins. Part VIII. Expression in Plants. Plant Cell Transfection by Electroporation, William B. Terzaghi and Anthony R. Cashmore. Transformation and Foreign Gene Expression in Plants Mediated by Microprojectile Bombardment, Henry Daniell. Transformation of the Cereals Using Agrobacterium, Jean Gould. Transformation of Microalgae Using Silicon Carbide Whiskers, Terri G. Dunahay, Sally A.