Heterologous Gene Expression in E.coli Methods and Protocols
This detailed volume provides a toolbox for designing constructs, tackling expression and solubility issues, handling membrane proteins and protein complexes, and exploring innovative engineering of E. coli. The topics are largely grouped under four parts
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Nicola A. Burgess-Brown Editor
Heterologous Gene Expression in E. coli Methods and Protocols
Methods
in
Molecular Biology
Series Editor John M. Walker School of Life and Medical Sciences University of Hertfordshire Hatfield, Hertfordshire, AL10 9AB, UK
For further volumes: http://www.springer.com/series/7651
Heterologous Gene Expression in E. coli Methods and Protocols
Edited by
Nicola A. Burgess-Brown SGC, Nuffield Department of Clinical Medicine, University of Oxford, Oxford, UK
Editor Nicola A. Burgess-Brown SGC Nuffield Department of Clinical Medicine University of Oxford Oxford, UK
ISSN 1064-3745 ISSN 1940-6029 (electronic) Methods in Molecular Biology ISBN 978-1-4939-6885-5 ISBN 978-1-4939-6887-9 (eBook) DOI 10.1007/978-1-4939-6887-9 Library of Congress Control Number: 2017934051 © Springer Science+Business Media LLC 2017 This work is subject to copyright. All rights are reserved by the Publisher, whether the whole or part of the material is concerned, specifically the rights of translation, reprinting, reuse of illustrations, recitation, broadcasting, reproduction on microfilms or in any other physical way, and transmission or information storage and retrieval, electronic adaptation, computer software, or by similar or dissimilar methodology now known or hereafter developed. The use of general descriptive names, registered names, trademarks, service marks, etc. in this publication does not imply, even in the absence of a specific statement, that such names are exempt from the relevant protective laws and regulations and therefore free for general use. The publisher, the authors and the editors are safe to assume that the advice and information in this book are believed to be true and accurate at the date of publication. Neither the publisher nor the authors or the editors give a warranty, express or implied, with respect to the material contained herein or for any errors or omissions that may have been made. The publisher remains neutral with regard to jurisdictional claims in published maps and institutional affiliations. Printed on acid-free paper This Humana Press imprint is published by Springer Nature The registered company is Springer Science+Business Media LLC The registered company address is: 233 Spring Street, New York, NY 10013, U.S.A.
Preface Heterologous gene expression in E. coli has been one of the most widely used methods for generating recombinant proteins for many scientific analyses and still remains the first choice for most laboratories around the world. The ease of use and low cost of production often lead researchers to initially attempt to express their proteins of interest in E. coli rather than opting for a eukaryotic host. Decades of development have seen the variety of methods for expressing genes in E. coli broaden, with improved media and optimized conditions for growth, a choice of promoter systems to regulate expression, fusion tags to aid solubility and purification, and E. coli host strains to accommodate more challenging or toxic proteins. Having work
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