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Gene Expression Technology : Volume 185 - John N. Abelson

Gene Expression Technology

Volume 185

By: John N. Abelson (Editor), Melvin I. Simon (Editor), David V. Goeddel (Editor)

Hardcover

Published: 1st May 1990
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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.

"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 Many of the contributors have been able to effectively communicate years worth of experience and unpublished observations in what often was a black magic science.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." --AMERICAN JOURNAL OF PHYSIOLOGY "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." --TRENDS IN GENETICS "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." --ANALYTICAL BIOCHEMISTRY "This work is essential for the experienced worker or novice who having cloned his gene wishes to express it." --BIOTECHNOLOGY & APPLIED BIOCHEMISTRY

Introduction
Systems for Heterologous Gene Expression
Expression in Escherichia coli
Expression of Heterologous Proteins in Escherichia coli
Vectors
Design and Construction of Expression Plasmid Vectors in Escherichia coli
Promoters
Promoter Recognition and mRNA Initiation by Escherichia coli Eo - 70
Use of Escherichia coli trp Promoter for Direct Expression of Proteins
Use of T7 RNA Polymerase to Direct Expression of Cloned Genes
Translation
High-Level Translation Initiation
Enhanced Translational Efficiency with Two-Cistron Expression System
Sequences within Ribosome Binding Site Affecting Messenger RNA Translatability and Method to Direct Ribosomes to Single Messenger RNA Species
Vector for Enhanced Translation of Foreign Genes in Escherichia coli
Proteases
Minimizing Proteolysis in Escherichia coli: Genetic Solutions
Gene Fusions
Gene Fusions for Purpose of Expression: An Introduction
Fusions to Staphylococcal Protein A
Expression as trpE Fusion
Secretion
Engineering Escherichia coli to Secrete Heterologous Gene Products
Protein Refolding
Refolding of Recombinant Proteins
Expression in Bacillus subtilis
Expression of Heterologous Genes in Bacillus subtilis
Intracellular Production
Regulated Promoter for High-Level Expression of Heterologous Genes in Bacillus subtilis
Secretion
System for Secretion of Heterologous Proteins in Bacillus subtilis
Inducible Expression of Regulatory Proteins
Inducible Expression of Regulatory Genes in Bacillus subtilis
Expression in Yeast
Heterologous Gene Expression in Yeast
Plasmid Vectors
Propagation and Expression of Cloned Genes in Yeast: 2-micro;Circle-Based Vectors
Manipulating Yeast Genome Using Plasmid Vectors
Inducible Expression Cassettes
Regulated GAL4 Expression Cassette Providing Controllable and High-Level Output from High-Copy Galactose Promoters in Yeast
Expression of Heterologous Proteins in Saccharomyces cerevisiae Using ADH2 Promoter
Induced Expression Using Yeast Coppe
Table of Contents provided by Publisher. All Rights Reserved.

ISBN: 9780121820862
ISBN-10: 0121820866
Series: Methods in Enzymology : Book 185
Audience: Professional
Format: Hardcover
Language: English
Number Of Pages: 681
Published: 1st May 1990
Publisher: Elsevier Science Publishing Co Inc
Country of Publication: US
Dimensions (cm): 22.9 x 15.2  x 3.18
Weight (kg): 1.12