Recombinant Gene Expression
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Recombinant Gene Expression : Reviews and Protocols

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Since newly created beings are often perceived as either wholly good or bad, the genetic alteration of living cells impacts directly on a symbolic meaning deeply imbedded in every culture. During the earlier years of gene expression research, te- nological applications were confined mainly to academic and industrial laboratories, and were perceived as highly beneficial since molecules that were previously unable to be separated or synthesized became accessible as therapeutic agents. Such were the success stories of hormones, antibodies, and vaccines produced in the bacterium Escherichia coli. Originally this bacterium gained fame among humans for being an unwanted host in the intestine, or worse yet, for being occasionally dangerous and pathogenic. H- ever, it was easily identified in contaminated waters during the 19th century, thus becoming a clear indicator of water pollution by human feces. Tamed, cultivated, and easily maintained in laboratories, its fast growth rate and metabolic capacity to adjust to changing environments fascinated the minds of scientists who studied and modeled such complex phenomena as growth, evolution, genetic exchange, infection, survival, adaptation, and further on-gene expression. Although at the lower end of the complexity scale, this microbe became a very successful model system and a key player in the fantastic revolution kindled by the birth of recombinant DNA technology.show more

Product details

  • Hardback | 508 pages
  • 157.5 x 231.1 x 33mm | 952.56g
  • Humana Press Inc.
  • Totowa, NJ, United States
  • English
  • Revised
  • 2nd ed. 2004
  • biography
  • 1588292622
  • 9781588292629

Review quote

From reviews of the first edition... "...particularly useful for teaching molecular biology laboratory courses." -Quarterly Review of Biology "...represents good value and will be a useful addition to the laboratory." -Microbiology Todayshow more

Back cover copy

In this updated and expanded second edition of an established classic, the editors have added critical reviews to a fresh collection of cutting-edge protocols for gene expression in bacteria, fungi, plants, plant cells, animals, and animal cells. The review articles survey new directions in recombinant gene expression research, technique, and application, and point the way to using recombinant gene expression for metabolic engineering and the production of nonprotein molecules. The readily reproducible protocols offer new host-vector systems, such as Antarctic bacteria and moderately halophytic bacteria, vectors for chromosomal editing and metabolic engineering, and information to aid in problem solving, strategy planning, and the promotion of protein folding. There are also novel heterologous gene expression schemes for cell-free systems, plant chloroplasts, microplates, hairy roots, and animal milk, and new developments in consolidated heterologous systems. The protocols follow the successful Methods in Molecular Biology series format, each one offering step-by-step laboratory instructions, an introduction outlining the principle behind the technique, lists of equipment and reagents, and tips on troubleshooting and avoiding known pitfalls. State-of-the-art and forward looking, Recombinant Gene Expression: Reviews and Protocols, Second Edition offers investigators seeking an overview of this critically important field not only the understanding, but also the tools needed to begin producing nonprotein products and altering the central metabolic pathways of cells to enhance heterologous gene expression."show more

Table of contents

Part I. General Issues About Recombinant Gene Expression Host Cell Compatibility in Protein Expression James J. Greene Production of Recombinant Proteins: Challenges and Solutions Laura A. Palomares, Sandino Estrada-Mondaca, and Octavio T. Ramirez Folding-Promoting Agents in Recombinant Protein Production Beatrix Fahnert Part II. Prokaryotes Back to Basics: pBR322 and Protein Expression Systems in E. coli Paulina Balbas and Francisco Bolivar a-Complementation Enabled T7 Expression Vectors and Their Use for the Expression of Recombinant Polypeptides for Protein Transduction Experiments Mikhail F. Alexeyev, Viktoriya V. Pastukh, Inna N. Shokolenko, and Glenn L. Wilson Expression of Recombinant Alkaline Phosphatase Conjugates in Escherichia coli Jean-Claude Boulain and Frederic Ducancel Overexpression of Chromosomal Genes in Escherichia coli Fernando Valle and Noemi Flores Chromosomal Expression of Foreign and Native Genes From Regulatable Promoters in Escherichia coli Lu Zhou, Ke Zhang, and Barry L. Wanner Plasmid Vectors for Marker-Free Chromosomal Insertion of Genetic Material in Escherichia coli Sylvie Le Borgne, Francisco Bolivar, and Guillermo Gosset Copy-Control pBAC/oriV Vectors for Genomic Cloning Jadwiga Wild and Waclaw Szybalski Copy-Control Tightly Regulated Expression Vectors Based on pBAC/oriV Jadwiga Wild and Waclaw Szybalski Cell-Free Protein Synthesis With Prokaryotic Combined Transcription-Translation James R. Swartz, Michael C. Jewett, and Kim A. Woodrow Genetic Tools for the Manipulation of Moderately Halophilic Bacteria of the Family Halomonadaceae Carmen Vargas and Joaquin J. Nieto Gene Transfer and Expression of Recombinant Proteins in Moderately Halophilic Bacteria Amalia S. Afendra, Carmen Vargas, JoaquinJ. Nieto, and Constantin Drainas Recombinant Protein Production in Antarctic Gram-Negative Bacteria Angela Duilio, Maria Luisa Tutino, and Gennaro Marino Part III. Fungi Recombinant Protein Production in Yeasts Danilo Porro and Diethard Mattanovich Controlled Expression of Homologous Genes by Genomic Promoter Replacement in the Yeast Saccharomyces cerevisiae Kevin J. Verstrepen and Johan M. Thevelein High-Throughput Expression in Microplate Format in Saccharomyces cerevisiae Caterina Holz and Christine Lang High-Throughput Expression in Microplate Format in Pichia pastoris Mewes Bottner and Christine Lang Multiple Gene Expression by Chromosomal Integration and CRE-loxP-Mediated Marker Recycling in Saccharomyces cerevisiae Bjorn Johansson and Barbel Hahn-Hagerdal Three Decades of Fungal Transformation: Key Concepts and Applications Vianey Olmedo-Monfil, Carlos Cortes-Penagos, and Alfredo Herrera-Estrella Three Decades of Fungal Transformation: Novel Technologies Sergio Casas-Flores, Teresa Rosales-Saavedra, and Alfredo Herrera-Estrella Part IV. Plants and Plant Cells Gene Transfer and Expression in Plants Argelia Lorence and Robert Verpoorte Production of Recombinant Proteins by Hairy Roots Cultured in Plastic Sleeve Bioreactors Fabricio Medina-Bolivar and Carole Cramer Engineering the Chloroplast Genome for Hyperexpression of Human Therapeutic Proteins and Vaccine Antigens Shashi Kumar and Henry Daniell New Selection Marker for Plant Transformation Barbara Leyman, Nelson Avonce, Matthew Ramon, Patrick Van Dijck, Johan M. Thevelein, and Gabriel Iturriaga Enhancer Detection and Gene Trapping as Tools for Functional Genomics in Plants Gerardo Acosta-Garcia, Daphne Autran, and Jean-Philippe Vielle-Calzada Part V. Animals andshow more

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