Enzyme Engineering [electronic resource] : Methods and Protocols / edited by James C. Samuelson.
- Totowa, NJ : Humana Press : Imprint: Humana Press, 2013.
- Methods in Molecular Biology, Methods and Protocols, 1064-3745 ; 978
Springer Protocols (Springer-12345)
Methods in Molecular Biology, Methods and Protocols, 1064-3745 ; 978
1 online resource (XI, 252 pages) : 52 illustrations, 19 illustrations in color.
- Life sciences.
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- Life Sciences.
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- text file PDF
- 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.
- A Tripartite Fusion System for the Selection of Protein Variants with Increased Stability in vivo
Determining Enzyme Kinetics via Isothermal Titration Calorimetry
GFP Reporter Screens for the Engineering of Amino Acid Degrading Enzymes from Libraries Expressed in Bacteria
Flow Cytometric Assays for Interrogating LAGLIDADG Homing Endonuclease DNA Binding and Cleavage Properties
TAL Effector Nuclease (TALEN) Engineering
In vitro Evolution of Enzymes
Residue-Specific Incorporation of Unnatural Amino Acids into Proteins In vitro and In vivo
Reconstructing Evolutionary Adaptive Paths for Protein Engineering
Oligonucleotide Recombination Enabled Site-Specific Mutagenesis in Bacteria
FX Cloning: A Versatile High-Throughput Cloning System for Characterization of Enzyme Variants
Use of Sulfolobus solfataricus PCNA Subunit Proteins to Direct the Assembly of Multimeric Enzyme Complexes
Gene Synthesis by Assembly of Deoxyuridine Containing Oligonucleotides
Protein Engineering: Single or Multiple Site-Directed Mutagenesis
Gene Assembly and Combinatorial Libraries in S. cerevisiae via Reiterative Recombination
Promiscuity-Based Enzyme Selection for Rational Directed Evolution Experiments
Rational Protein Sequence Diversification by Multi-Codon Scanning Mutagenesis
Screening Libraries for Improved Solubility: Using E. coli Dihydrofolate Reductase as a Reporter
In Vitro Directed Evolution of Enzymes Expressed by E. coli in Micro-Titre Plates.
- Samuelson, James C. editor.
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- 10.1007/978-1-62703-293-3 doi
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- Restricted for use by site license.
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