Protein Engineering Protocols

Comprehensive Guide to Methods at Every Stage of the Engineering Process

Protein Engineering Protocols considers the general, but not mutually exclusive, strategies for protein engineering. The first is known as rational design, in which the scientist uses detailed knowledge of the structure and function of the protein to make desired changes. The second strategy is known as directed evolution. This volume provides a comprehensive guide to the methods used at every stage of the engineering process. It combines a thorough theoretical foundation with detailed protocols and will be invaluable to all research workers in the area, from graduate students to senior investigators.

Inhalt
Design and Computational Strategies for Protein Engineering.- Combinatorial Protein Design Strategies Using Computational Methods.- Global Incorporation of Unnatural Amino Acids in Escherichia coli.- Considerations in the Design and Optimization of Coiled Coil Structures.- Calcium Indicators Based on Calmodulin-Fluorescent Protein Fusions.- Design and Synthesis of Artificial Zinc Finger Proteins.- Monobodies.- Engineering Site-Specific Endonucleases.- Evolutionary Strategies for Protein Engineering.- Protein Library Design and Screening.- Protein Design by Binary Patterning of Polar and Nonpolar Amino Acids.- Versatile DNA Fragmentation and Directed Evolution With Nucleotide Exchange and Excision Technology.- Degenerate Oligonucleotide Gene Shuffling.- M13 Bacteriophage Coat Proteins Engineered for Improved Phage Display.- Ribosome-Inactivation Display System.- Compartmentalized Self-Replication.- Synthesis of Degenerated Libraries of the Ras-Binding Domain of Raf and Rapid Selection of Fast-Folding and Stable Clones With the Dihydrofolate Reductase Protein Fragment Complementation Assay.- A General Method of Terminal Truncation, Evolution, and Re-Elongation to Generate Enzymes of Enhanced Stability.