Gene Expression Systems: Using Nature for the Art of Expression offers detailed information on a wide variety of gene expression systems from an array of organisms. It describes several different types of expression systems including transient, stable, viral, and transgenic systems. Each chapter is written by a leader in the field. The book includes timelines and examples for each expression system, and provides an overview of the future of recombinant protein expression. Key Features * Provides detailed information on expression systems * Covers a variety of promoters and host organisms enabling researchers to tailor protocols to their specific needs * Includes timelines and examples * Compares pros and cons of each method
E-Book Content
Gene Expression Systems
This Page Intentionally Left Blank
Gene Expression Systems Using Nature for the Art of Expression
Edited by Joseph M. Fernandez and James P. Hoeffler Invitrogen Corporation Carlsbad, California
ACADEMIC PRESS San Diego
London Boston New York Sydney Tokyo Toronto
This book is printed on acid-free paper. ( ~ Copyright 9 1999 by ACADEMIC PRESS All Rights Reserved. No part of this publication may be reproduced or transmitted in any form or by any means, electronic or mechanical, including photocopy, recording, or any information storage and retrieval system, without permission in writing from the publisher.
Academic Press a division of Harcourt Brace & Company 525 B Street, Suite 1900, San Diego, California 92101-4495, USA http://www.apnet.com
Academic Press 24-28 Oval Road, London NW 1 7DX, UK http://www.hbuk.co.uk/ap/ Library of Congress Catalog Card Number: 98-85621 International Standard Book Number: 0-12-253840-4 PRINTED IN THE UNITED STATES OF AMER/CA 98 99 00 01 02 03 MM 9 8 7 6
5
4
3
2
1
CONTENTS
Contributors
xi
Introduction Joseph M. Fernandez and James P. Hoeffler The Art of Expression 1 Purpose of This Book 2 Selecting a Suitable Expression System: Considerations 3 Genomics and the Future of Protein Expression Systems 5
Section I PROKARYOTIC EXPRESSION SYSTEMS 1. Gene Expression Systems Based on Bacteriophage T7 R N A Polymerase Russell Durbin Introduction 10 T7 RNAP in Its Natural Habitat 11 Biochemistry of T7 RNAP 14 T7-Based Gene Expression Systems 24 Conclusions 36 References 37 2. Expression Vectors Employing the trc Promoter Jfirgen Brosius Introduction 46 Vectors for trc-Driven Expression 51 Additional Hosts for trc-Driven Expression 57 Outlook for Further Improvements of Expression Vectors Employing the trc Promoter 58 References 59
vi
Contents
3. Bacillus Expression" A Gram-Positive Model Eugenio Ferrari and Brian Miller Introduction 66 Bacilli Used as Industrial Production Organisms Genetic Manipulation in Bacilli 71 Plasmid Vectors 75 Transcriptional Regulation 77 Conclusions 81 Appendix 82 References 89
67
4. araB Expression System in Escherichia coli Marc Better
Introduction 95 The araB Promoter and How It Works 97 Use of the ara System for Expression of Recombinant Products 99 Conclusions 104 References 106
Section II EUKARYOTIC EXPRESSION SYSTEMS 5. Adenoviral Vectors for Protein Expression Dan J. Von Seggern and Glen R. Nemerow Adenovirus Biology 112 Adaptation of Adenovirus as a Gene Transfer Vector 117 Construction of Adenovirus Expression Vectors 119 Applications of Adenoviral Vectors in Gene Expression 128 Considerations in the Use of Adenovirus Expression Vectors 138 Advantages and Disadvantages of Adenoviral Vectors 141 Future Directions 143 Conclusions 144 References 145
Contents
vii
6. Expression in the Methylotrophic Yeast Pichia pastoris James M. Cregg Introduction 158 Background Informatio