Enzyme Engineering

Methods and Protocols

Specificaties
Paperback, blz. | Engels
Humana Press | e druk, 2016
ISBN13: 9781493962877
Rubricering
Humana Press e druk, 2016 9781493962877
Onderdeel van serie Methods in Molecular Biology
Verwachte levertijd ongeveer 9 werkdagen

Samenvatting

Whether the pursuit is commercially motivated or purely academic, engineering a novel biological catalyst is an enticing challenge. High-resolution protein structure analysis allows for rational alteration of enzyme function, yet many useful enzyme variants are the product of well-designed selection schemes or screening strategies. Enzyme Engineering: Methods and Protocols provides guidance to investigators wishing to create enzyme variants with desired properties. This detailed volume covers such topics as a simple method for generating site-specific mutations within bacterial chromosomes. It also highlights the engineering of two difference types of rare-cutting endonucleases that show great potential in gene therapy applications: The newest development is the emergence of TAL effector nucleases or TALENs. Chapters describe newly developed technologies in sufficient detail so that each method can be practiced in a standard molecular biology laboratory. Written in the successful Methods in Molecular Biology™ series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols, and notes on troubleshooting and avoiding known pitfalls.

 

Authoritative and easily accessible Enzyme Engineering: Methods and Protocols will be valuable for scientists with a budding interest in protein engineering as well as veterans looking for new approaches to apply in established discovery programs.

Specificaties

ISBN13:9781493962877
Taal:Engels
Bindwijze:paperback
Uitgever:Humana Press

Inhoudsopgave

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

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