Directed Enzyme Evolution: Screening and Selection Methods

Part I. Genetic Selections
Genetic Complementation Protocols
Jessica L. Sneeden and Lawrence A. Loeb
Use of Pol I-Deficient E. coli for Functional Complementation of DNA Polymerase
Manel Camps and Lawrence A. Loeb
Selection of Novel Eukaryotic DNA Polymerases by Mutagenesis and Genetic Complementation of Yeast
Ranga N. Venkatesan and Lawrence A. Loeb
Autogene Selections
Jijumon Chelliserrykattil and Andrew D. Ellington
Selection for Soluble Proteins via Fusion with Chloramphenicol Acetyltransferase
Volker Sieber
Proside: A Phage-Based Method for Selecting Thermostable Proteins
Andreas Martin, Franz X. Schmid, and Volker Sieber
Minimization of Proteins by Random Fragmentation and Selection
Gary W. Rudgers and Timothy Palzkill
Part II. Screens for Enzymes
Evaluating a Screen and Analysis of Mutant Libraries
Oriana Salazar and Lianhong Sun
Screening Mutant Libraries in Saccharomyces cerevisiae
Thomas Bulter, Volker Sieber, and Miguel Alcalde
Solid-Phase Screening Using Digital Image Analysis
Alexander V. Tobias and John M. Joern
Screening for Thermostability
Patrick C. Cirino and Radu Georgescu
High-Throughput Screening of Mutant a-Amylase Libraries for Increased Activity at 129°C Holger Berk and Robert J. Lebbink
High-Throughput Carbon Monoxide Binding Assay for Cytochromes P450
Christopher R. Otey
High-Throughput Screen for Aromatic Hydroxylation
Christopher R. Otey and John M. Joern
Colorimetric Screen for Aliphatic Hydroxylation by Cytochrome P450 Using p-Nitrophenyl-Substituted Alkanes
Edgardo T. Farinas
High-Throughput Screens Based on NAD(P)H Depletion
Anton Glieder and Peter Meinhold
High-Throughput Tetramethylbenzidine (TMB) Screen for Peroxidases
Radu Georgescu
Screenfor Oxidases by Detection of Hydrogen Peroxide with Horseradish Peroxidase
Lianhong Sun and Makoto Yagasaki
Colorimetric Dehydrogenase Screen Based on NAD(P)H Generation
Kimberly M. Mayer
Colorimetric Assays for Screening Laccases
Miguel Alcalde and Thomas Bulter
pH Sensing Agar Plate Assays for Esterolytic Enzyme Activity
Karl E. Griswold
A pH-Indicator-Based Screen for Hydrolytic Haloalkane Dehalogenase
Huimin Zhao
Detection of Aromatic a-Hydroxyketones with Tetrazolium Salts
Michael Breuer and Bernhard Hauer
Selection of Heat-Stable Clostridium cellulovorans Cellulases After In Vitro Recombination
Koichiro Murashima and Roy H. Doi
Screening and Selection Strategies for Disulfide Isomerase Activity
Ronald Lafond, Xiaoming Zhan, and George Georgiou
An Overview of High-Throughput Screening Systems for Enantioselective Enzymatic Transformations
Manfred T. Reetz
Select Protocols of High-Throughput ee-Screening Systems for Assaying Enantioselective Enzymes
Manfred T. Reetz
Directed Evolution of the Substrate Specificities of a Site-Specific Recombinase and an Aminoacyl-tRNA Synthetase Using Fluorescence-Activated Cell Sorting (FACS)
Stephen W. Santoro and Peter G. Schultz
Calmodulin-Tagged Phage and Two-Filter Sandwich Assays for the Identification of Enzymatic Activities
Christian Heinis, Julian Bertschinger, and Dario Neri
High-Throughput FACS Method for Directed Evolution of Substrate Specificity
Mark J. Olsen, Jongsik Gam, Brent L. Iverson, and George Georgiou
Improving Protein Folding Efficiency by Directed Evolution Using the GFP Folding Reporter
Geoffrey S. Waldo
Index