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  • Book
  • © 2003

Directed Enzyme Evolution

Screening and Selection Methods

Editors:

  1. Frances H. Arnold

    1. California Institute of Technology, Pasadena

    View editor publications

    You can also search for this editor in PubMed   Google Scholar

  2. George Georgiou

    1. University of Texas at Austin, Austin

    View editor publications

    You can also search for this editor in PubMed   Google Scholar

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

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Table of contents (31 protocols)

  1. Front Matter

    Pages i-xv
    PDF

  2. Genetic Selections

    1. Front Matter

      Pages 1-1
      PDF

    2. Genetic Complementation Protocols

      • Jessica L. Sneeden, Lawrence A. Loeb
      Pages 3-10

    5. Autogene Selections

      • Jijumon Chelliserrykattil, Andrew D. Ellington
      Pages 27-43

    7. Proside

      • Andreas Martin, Franz X. Schmid, Volker Sieber
      Pages 57-70

    8. Minimization of Proteins by Random Fragmentation and Selection

      • Gary W. Rudgers, Timothy Palzkill
      Pages 71-81

  3. Screens for Enzymes

    1. Front Matter

      Pages 83-83
      PDF

    2. Evaluating a Screen and Analysis of Mutant Libraries

      • Oriana Salazar, Lianhong Sun
      Pages 85-97

    3. Screening Mutant Libraries in Saccharomyces cerevisiae

      • Thomas Bulter, Volker Sieber, Miguel Alcalde
      Pages 99-107

    4. Solid-Phase Screening Using Digital Image Analysis

      • Alexander V. Tobias, John M. Joern
      Pages 109-115

    5. Screening for Thermostability

      • Patrick C. Cirino, Radu Georgescu
      Pages 117-125

    8. High-Throughput Screen for Aromatic Hydroxylation

      • Christopher R. Otey, John M. Joern
      Pages 141-148

    10. High-Throughput Screens Based on NAD(P)H Depletion

      • Anton Glieder, Peter Meinhold
      Pages 157-170
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About this book

Directed evolution comprises two distinct steps that are typically applied in an iterative fashion: (1) generating molecular diversity and (2) finding among the ensemble of mutant sequences those proteins that perform the desired fu- tion according to the specified criteria. In many ways, the second step is the most challenging. No matter how cleverly designed or diverse the starting library, without an effective screening strategy the ability to isolate useful clones is severely diminished. The best screens are (1) high throughput, to increase the likelihood that useful clones will be found; (2) sufficiently sen- tive (i. e. , good signal to noise) to allow the isolation of lower activity clones early in evolution; (3) sufficiently reproducible to allow one to find small improvements; (4) robust, which means that the signal afforded by active clones is not dependent on difficult-to-control environmental variables; and, most importantly, (5) sensitive to the desired function. Regarding this last point, almost anyone who has attempted a directed evolution experiment has learned firsthand the truth of the dictum “you get what you screen for. ” The protocols in Directed Enzyme Evolution describe a series of detailed p- cedures of proven utility for directed evolution purposes. The volume begins with several selection strategies for enzyme evolution and continues with assay methods that can be used to screen enzyme libraries. Genetic selections offer the advantage that functional proteins can be isolated from very large libraries s- ply by growing a population of cells under selective conditions.
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Reviews

"...covers a considerable number of protocols for a broad range of enzymes...very useful..." - ChemBioChem
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Editors and Affiliations

  • California Institute of Technology, Pasadena

    Frances H. Arnold

  • University of Texas at Austin, Austin

    George Georgiou

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Bibliographic Information

  • Book Title: Directed Enzyme Evolution

  • Book Subtitle: Screening and Selection Methods

  • Editors: Frances H. Arnold, George Georgiou

  • Series Title: Methods in Molecular Biology

  • DOI: https://doi.org/10.1385/1592593968

  • Publisher: Humana Totowa, NJ

  • eBook Packages: Springer Protocols

  • Copyright Information: Humana Press 2003

  • Hardcover ISBN: 978-1-58829-286-5Published: 16 May 2003

  • Softcover ISBN: 978-1-61737-472-2Published: 10 November 2010

  • eBook ISBN: 978-1-59259-396-5Published: 02 February 2008

  • Series ISSN: 1064-3745

  • Series E-ISSN: 1940-6029

  • Edition Number: 1

  • Number of Pages: XV, 383

  • Topics: Cell Biology

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Buy it now

eBook USD 129.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book USD 169.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book USD 169.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Other ways to access

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