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.
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Product details

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

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(TM) 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.
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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 Boettner and Christine Lang

Multiple Gene Expression by Chromosomal Integration and CRE-loxP-Mediated Marker Recycling in Saccharomyces cerevisiae
Bjoern 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 and
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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 Today
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