Enzyme Engineering : Methods and Protocols

1. A Tripartite Fusion System for the Selection of Protein Variants with Increased Stability in vivo Linda Foit and James Bardwell 2. Determining Enzyme Kinetics via Isothermal Titration Calorimetry Neil A. Demarse, Marie C. Killian, Lee D. Hansen, and Colette F. Quinn 3. GFP Reporter Screens for the Engineering of Amino Acid Degrading Enzymes from Libraries Expressed in Bacteria Olga Paley, Giulia Agnello, Jason Cantor, Tae Hyun Yoo, George Georgiou, and Everett Stone 4. Flow Cytometric Assays for Interrogating LAGLIDADG Homing Endonuclease DNA Binding and Cleavage Properties Sarah K. Baxter, Abigail R. Lambert, Andrew M. Scharenberg, and Jordan Jarjour 5. TAL Effector Nuclease (TALEN) Engineering Ting Li and Bing Yang 6. In vitro Evolution of Enzymes Misha V. Golynskiy, John C. Haugner III, Aleardo Morelli, Dana Morrone, and Burckhard Seelig 7. Residue-Specific Incorporation of Unnatural Amino Acids into Proteins In vitro and In vivo Amrita Signh-Blom, Randall A. Hughes, and Andrew. D. Ellington 8. Reconstructing Evolutionary Adaptive Paths for Protein Engineering Megan F. Cole, Vanessa E. Cox, Kelsey L. Gratton, and Eric A. Gaucher 9. Oligonucleotide Recombination Enabled Site-Specific Mutagenesis in Bacteria Bryan M. Swingle 10. FX Cloning: A Versatile High-Throughput Cloning System for Characterization of Enzyme Variants Eric R. Geertsma 11. Use of Sulfolobus solfataricus PCNA Subunit Proteins to Direct the Assembly of Multimeric Enzyme Complexes Hidehiko Hirakawa and Teruyuki Nagamune 12. Gene Synthesis by Assembly of Deoxyuridine Containing Oligonucleotides Romualdas Vaisvila and Jurate Bitinaite 13. Protein Engineering: Single or Multiple Site-Directed Mutagenesis Pei-Chung Hsieh and Romualdas Vaisvila 14. Gene Assembly and Combinatorial Libraries in S. cerevisiae via Reiterative Recombination Nili Ostrov, Laura M. Wingler, and Virginia Cornish 15. Promiscuity-Based Enzyme Selection for Rational Directed Evolution Experiments Sandeep Chakraborty, Renu Minda, Lipika Salaye, Abhaya M. dandekar, Swapan K. Bhattacharjee, and Basuthkar J. Rao 16. Rational Protein Sequence Diversification by Multi-Codon Scanning Mutagenesis Jia Liu and T. Ashton Cropp 17. Screening Libraries for Improved Solubility: Using E. coli Dihydrofolate Reductase as a Reporter Jian-Wei Liu and David Ollis 18. In Vitro Directed Evolution of Enzymes Expressed by E. coli in Micro-Titre Plates Bradley J. Stevenson, Sylvia H.C. Yip, and David L. Ollis