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020 _a9781627032933
024 7 _a10.1007/978-1-62703-293-3
_2doi
040 _aES-MaUEC
_bspa
_cES-MaUEC
245 1 0 _aEnzyme Engineering
_bMethods and Protocols
_cedited by James C. Samuelson.
250 _a1st edition 2013
264 1 _aTotowa, NJ
_bHumana Press
_c2013
300 _a1 recurso en línea (XI, 252 páginas)
_b52 ilustraciones, 19 ilustraciones a color
336 _atexto
_btxt
_2rdacontent
337 _aelectrónico
_bc
_2rdamedia
338 _arecurso electrónico
_bcr
_2rdacarrier
347 _aarchivo de texto
_bPDF
490 0 _aMethods in Molecular Biology
_x1940-6029
_v978
505 0 _aA 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.
520 _aWhether 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.
700 1 _aSamuelson, James C
_eeditor literario
_4edt
_4http://id.loc.gov/vocabulary/relators/edt
776 0 8 _iPrinted edition:
_z9781627032940
776 0 8 _iPrinted edition:
_z9781627032926
776 0 8 _iPrinted edition:
_z9781493962877
856 4 0 _uhttps://go.openathens.net/redirector/universidadeuropea.es?url=https://doi.org/10.1007/978-1-62703-293-3
_z(usuarios Universidad Europea de Valencia)
942 _2lcc
_cLE
988 _aSpringer_Protocols_2013
999 _c234642
_d234642