Directed Enzyme Evolution / edited by Frances H. Arnold. -- (Methods in Molecular Biology ; v. 230). -- Human Press, 2003

索书号: 58.17/M592/v.230

Contents:

1.Genetic Complemetation Protocols  3

2.Use of Pol I-Deficient E. coli for Functional Complementation of DNA Polymerase  11

3.Selection of Novel Eukaryotic DNA Polymerases by Mutagenesis and Genetic Complementation of Yeast  19

4.Autogene Selections  27

5.Selection for Soluble Proteins via Fusion with Shloramphenicol Acetyltransferase  45

6.Proside: A Phage-Based Method for Selecting Thermostable Proteins  57

7.Minimization of Proteins by Random Fragmentation and Selection  71

8.Evaluating a Screen and Analysis of Mutant Libraries  85

9.Screening Mutant Libraries in Saccharomyces cerevisiae  99

10.Soid-phase Screening Using Digital Image Analysis  109

11.Screening for Thermostability  117

12.High-Throughput Screening of Mutant α -Amylase Libraries for Increased Activity at 129℃   127

13.High-Throughput Carbon Monoxide Binding Assay for Cytochromes P450  137

14.High-Throughput Screen for Aromatic Hydroxylation  141

15.Colorimetric Screen for Aliphatic Hydroxylation by Cytochrome P450 Using P-Nitrophenyl-Substituted Alkanes  149

16.High-Throughput Screens Based on NAD(P)H Depletion  157

17.High-Throughput Tetramethylbenzidine(TMB) Screen for Peroxidases  171

18.Screen for Oxidases by Detection of Hydrogen Peroxide with Horseradish Peroxidase  177

19.Colorimetric Dehydrogenase Screen Based on NAD(P)H Generation  183

20.Colorimetric Assays for Screening Laccases  193

21.pH Sensing Agar Plate Assays for Esterolytic Enzyme Activity  203

22.A pH-Induicator-Based Screen for Hydrolytic Haloalkane Dehalogenase  213

23. ... ...

31.Improving Protein Folding Efficiency by Directed Evolution Using the GFP Folding Reporter  343