Directed Evolution Library Creation

Methods and Protocols

  • Frances H. Arnold
  • George Georgiou

Part of the Methods in Molecular Biology™ book series (MIMB, volume 231)

Table of contents

  1. Front Matter
    Pages i-x
  2. Mutagenesis and Recombination Methods

    1. Front Matter
      Pages 1-1
    2. Patrick C. Cirino, Kimberly M. Mayer, Daisuke Umeno
      Pages 3-9
    3. Alexander V. Tobias
      Pages 11-16
    4. Thomas Bulter, Miguel Alcalde
      Pages 17-22
    5. Paul H. Bessette, Marco A. Mena, Annalee W. Nguyen, Patrick S. Daugherty
      Pages 29-37
    6. Annalee W. Nguyen, Patrick S. Daugherty
      Pages 39-44
    7. Olga Selifonova, Volker Schellenberger
      Pages 45-52
    8. Hiroshi Murakami, Takahiro Hohsaka, Masahiko Sisido
      Pages 53-64
    9. Jessica L. Sneeden, Lawrence A. Loeb
      Pages 65-73
    10. Radu Georgescu, Geethani Bandara, Lianhong Sun
      Pages 75-83
    11. John M. Joern
      Pages 85-89
    12. Wenjuan Zha, Tongbo Zhu, Huimin Zhao
      Pages 91-97
    13. Olga Esteban, Ryan D. Woodyer, Huimin Zhao
      Pages 99-104
    14. Anna Marie Aguinaldo, Frances H. Arnold
      Pages 105-110
    15. Wayne M. Coco
      Pages 111-127
    16. Marc Ostermeier, Stefan Lutz
      Pages 129-141
    17. Stefan Lutz, Marc Ostermeier
      Pages 143-151
    18. Andrew K. Udit, Jonathan J. Silberg, Volker Sieber
      Pages 153-163

About this book

Introduction

Directed protein evolution-a powerful technique for the discovery of new enzymes and therapeutic proteins-has matured to the point that it is now the centerpiece of enzyme engineering, where it has catalyzed the development of numerous experimental methods. In Directed Evolution Library Creation: Methods and Protocols, seasoned practitioners from many leading laboratories share their best experimental protocols for the generation of molecular diversity. Described in step-by-step detail to ensure experimental success, these protocols include readily reproducible methods for random mutagenesis of entire genes or segments of genes, for homologous and nonhomologous recombination, and for constructing in vivo libraries in bacteria and yeast. In addition to the various protocols for creating libraries, this volume also describes ways to analyze libraries, particularly those made by recombination. An accompanying volume, Directed Enzyme Evolution: Screening and Selection Methods (ISBN: 1-58829-286-X), is devoted entirely to selection and screening methods that can be applied to the directed evolution of enzymes.
Taken together, Directed Evolution Library Creation: Methods and Protocols and Directed Enzyme Evolution: Screening and Selection Methods capture for newcomers and more experienced investigators alike all the key methods for using directed protein evolution to better understand protein structure-function relationships, to discover new enzymes and therapeutic proteins, and to design new assays suitable for specific applications.

Editors and affiliations

  • Frances H. Arnold
    • 1
  • George Georgiou
    • 2
  1. 1.California Institute of TechnologyPasadena
  2. 2.University of Texas at AustinAustin

Bibliographic information

  • DOI https://doi.org/10.1385/159259395X
  • Copyright Information Humana Press 2003
  • Publisher Name Humana Press
  • eBook Packages Springer Protocols
  • Print ISBN 978-1-58829-285-8
  • Online ISBN 978-1-59259-395-8
  • Series Print ISSN 1064-3745
  • Series Online ISSN 1940-6029
  • About this book