Heterologous Gene Expression in E.coli Methods and Protocols
Protein expression in a heterologous host is a cornerstone of biomedical research and of the biotechnology industry. Despite the advanced state of protein expression technology improvements are still needed. For example, membrane proteins constitute a sig
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MO L E C U L A R BI O L O G Y
Series Editor John M. Walker School of Life Sciences University of Hertfordshire Hatfield, Hertfordshire, AL10 9AB, UK
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Heterologous Gene Expression in E. coli Methods and Protocols
Edited by
Thomas C. Evans, Jr. New England Biolabs, Inc., Ipswich, MA, USA
Ming-Qun Xu New England Biolabs, Inc., Ipswich, MA, USA
Editors Thomas C. Evans, Jr., PhD DNA Enzymes Division Head New England Biolabs, Inc. Ipswich, MA 01938, USA
Ming-Qun Xu, PhD Senior Scientist New England Biolabs, Inc. Ipswich, MA 01938, USA
ISSN 1064-3745 e-ISSN 1940-6029 ISBN 978-1-61737-966-6 e-ISBN 978-1-61737-967-3 DOI 10.1007/978-1-61737-967-3 Springer New York Dordrecht Heidelberg London © Springer Science+Business Media, LLC 2011 All rights reserved. This work may not be translated or copied in whole or in part without the written permission of the publisher (Humana Press, c/o Springer Science+Business Media, LLC, 233 Spring Street, New York, NY 10013, USA), except for brief excerpts in connection with reviews or scholarly analysis. Use in connection with any form of information storage and retrieval, electronic adaptation, computer software, or by similar or dissimilar methodology now known or hereafter developed is forbidden. The use in this publication of trade names, trademarks, service marks, and similar terms, even if they are not identified as such, is not to be taken as an expression of opinion as to whether or not they are subject to proprietary rights. While the advice and information in this book are believed to be true and accurate at the date of going to press, neither the authors nor the editors nor the publisher can accept any legal responsibility for any errors or omissions that may be made. The publisher makes no warranty, express or implied, with respect to the material contained herein. Printed on acid-free paper Humana Press is part of Springer Science+Business Media (www.springer.com)
Preface As we move solidly into the 21st century, it is amazing to see that molecular biology and its related disciplines are advancing at an ever-increasing pace. Today, it is commonplace to use bioinformatic techniques to identify a gene encoding a protein of interest and then pay a company to synthesize it, codon optimized, and receive it already inserted into the expression vector of choice. This is all the more incredible when one considers that less than 100 years ago scientists were still trying to identify the substance responsible for inherited traits, a little over 50 years ago the first DNA polymerase was discovered, and only about 40 years ago the first restriction enzyme was purified. It was the 1970s and 1980s that saw the expansive application of the prior basic research findings lead to the commonplace expression of proteins in a heterologous host. During this time recombinant DNA was first produced and shown to be stably maintained and replicated in Escherichia coli. The first recombinant protein, E. coli
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