Directed Enzyme Evolution

Screening and Selection Methods

  • Frances H. Arnold
  • George Georgiou

Part of the Methods in Molecular Biology™ book series (MIMB, volume 230)

Table of contents

  1. Front Matter
    Pages i-xv
  2. Genetic Selections

    1. Front Matter
      Pages 1-1
    2. Jessica L. Sneeden, Lawrence A. Loeb
      Pages 3-10
    3. Jijumon Chelliserrykattil, Andrew D. Ellington
      Pages 27-43
    4. Andreas Martin, Franz X. Schmid, Volker Sieber
      Pages 57-70
    5. Gary W. Rudgers, Timothy Palzkill
      Pages 71-81
  3. Screens for Enzymes

    1. Front Matter
      Pages 83-83
    2. Oriana Salazar, Lianhong Sun
      Pages 85-97
    3. Thomas Bulter, Volker Sieber, Miguel Alcalde
      Pages 99-107
    4. Alexander V. Tobias, John M. Joern
      Pages 109-115
    5. Patrick C. Cirino, Radu Georgescu
      Pages 117-125
    6. Christopher R. Otey, John M. Joern
      Pages 141-148
    7. Anton Glieder, Peter Meinhold
      Pages 157-170

About this book

Introduction

Directed evolution, the application of evolutionary design to enzyme engineering, requires effective screening strategies to isolate those proteins that perform a desired function from the libraries generated by the techniques. In Directed Enzyme Evolution: Screening and Selection Methods, seasoned practitioners from many leading laboratories describe their leading and readily reproducible screening strategies for isolating useful clones. These techniques have been optimized for sensitivity, high throughput, and robustness, and are of proven utility for directed evolution purposes. The assays presented use a variety of techniques, including genetic complementation, microtiter plates, solid-phase screens with colorimetric substrates, and flow cytometric screens. There are also representative examples of how phage libraries may be interrogated for enzymatic activity. Each protocol contains detailed step-by-step instructions and many notes on how best to deal with the problems that may occur. An accompanying volume, Directed Evolution Library Creation: Methods and Protocols (ISBN 1-58829-285-1), describes readily reproducible methods for the creation of mutated DNA molecules and DNA libraries.
Taken together, Directed Enzyme Evolution: Screening and Selection Methods and Directed Evolution Library Creation: Methods and Protocols 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/10.1385/1592593968
  • Copyright Information Humana Press 2003
  • Publisher Name Humana Press
  • eBook Packages Springer Protocols
  • Print ISBN 978-1-58829-286-5
  • Online ISBN 978-1-59259-396-5
  • Series Print ISSN 1064-3745
  • Series Online ISSN 1940-6029
  • About this book