Two Hybrid Technologies Methods and Protocols
Protein–protein interactions (PPIs) are strongly predictive of functional relationships among proteins in virtually all processes that take place in the living cell. Therefore, the comprehensive exploration of interactome networks is one of the major goal
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IN
MOLECULAR BIOLOGY™
Series Editor John M. Walker School of Life Sciences University of Hertfordshire Hatfield, Hertfordshire, AL10 9AB, UK
For further volumes: http://www.springer.com/series/7651
Two Hybrid Technologies Methods and Protocols
Edited by
Bernhard Suter Max-Delbrück-Centrum für Molekulare Medizin, Quintara Biosciences, Albany, CA, USA
Erich E. Wanker Max-Delbrück-Centrum für Molekulare Medizin, Berlin-Buch, Germany
Editors Bernhard Suter Max-Delbrück-Centrum für Molekulare Medizin Quintara Biosciences Albany, CA, USA [email protected]
Erich E. Wanker Max-Delbrück-Centrum für Molekulare Medizin Berlin-Buch, Germany [email protected]
ISSN 1064-3745 e-ISSN 1940-6029 ISBN 978-1-61779-454-4 e-ISBN 978-1-61779-455-1 DOI 10.1007/978-1-61779-455-1 Springer New York Dordrecht Heidelberg London Library of Congress Control Number: 2011940814 © Springer Science+Business Media, LLC 2012 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. Printed on acid-free paper Humana Press is part of Springer Science+Business Media (www.springer.com)
Preface Protein–protein interactions (PPIs) are strongly predictive of functional relationships among proteins in virtually all processes that take place in the living cell. Therefore, the comprehensive exploration of interactome networks is one of the major goals in systems biology. The development of “interactomics” as a field is largely driven by the development of innovative technologies and strategies for efficient screening, scoring, and validation of PPIs. The aim of this book is to provide a compendium of state-of-the art-protocols for the investigation of binary PPIs with the classical yeast two-hybrid (Y2H) approach, Y2H variants, and other in vivo methods for PPI mapping. Given the broad range of methodologies currently available, biochemical approaches like proteome-wide co-immunoprecipitation, and other in vitro and in vivo methodologies are not to be considered here. It needs to be emphasized, however, that alternative methods are very important for the complementation and validation of Y2H screens. The book is structured into two sections. The first gives a survey of protocols that are currently employed for Y2H high-throughput screens by different expert labs in the field. Rather than detailing the principles of screening, which have been described previ
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