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020 _a9781592593958
024 7 _a10.1385/159259395X
_2doi
040 _aES-MaUEC
_bspa
_cES-MaUEC
245 1 0 _aDirected Evolution Library Creation
_bMethods and Protocols
_cedited by Frances H. Arnold, George Georgiou.
250 _a1st edition 2003
264 1 _aTotowa, NJ
_bHumana Press
_c2003
300 _a1 recurso en línea (X, 226 páginas)
_b
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
_v231
505 0 _aMutagenesis and Recombination Methods -- Generating Mutant Libraries Using Error-Prone PCR -- Preparing Libraries in Escherichia coli -- Preparing Libraries in Saccharomyces cerevisiae -- Creating Random Mutagenesis Libraries by Megaprimer PCR of Whole Plasmid (MEGAWHOP) -- Construction of Designed Protein Libraries Using Gene Assembly Mutagenesis -- Production of Randomly Mutated Plasmid Libraries Using Mutator Strains -- Evolution of Microorganisms Using Mutator Plasmids -- Random Insertion and Deletion Mutagenesis -- Random Oligonucleotide Mutagenesis -- Saturation Mutagenesis -- DNA Shuffling -- Family Shuffling with Single-Stranded DNA -- In Vitro DNA Recombination by Random Priming -- Staggered Extension Process (StEP) In Vitro Recombination -- RACHITT -- The Creation of ITCHY Hybrid Protein Libraries -- Preparation of SCRATCHY Hybrid Protein Libraries -- Sequence Homology-Independent Protein Recombination (SHIPREC) -- Producing Chimeric Genes by CLERY -- Analysis of Library Diversity -- Analysis of Shuffled Libraries by Oligonucleotide Probe Hybridization -- Sequence Mapping of Combinatorial Libraries on Macro- or Microarrays -- Sequence Mapping of Combinatorial Libraries on Macro- and Microarrays.
520 _aDirected protein evolution-a powerful technique for the discovery of new enzymes and therapeutic proteins-has matured to the point that it is now the centerpiece of enzyme engineering, where it has catalyzed the development of numerous experimental methods. In Directed Evolution Library Creation: Methods and Protocols, seasoned practitioners from many leading laboratories share their best experimental protocols for the generation of molecular diversity. Described in step-by-step detail to ensure experimental success, these protocols include readily reproducible methods for random mutagenesis of entire genes or segments of genes, for homologous and nonhomologous recombination, and for constructing in vivo libraries in bacteria and yeast. In addition to the various protocols for creating libraries, this volume also describes ways to analyze libraries, particularly those made by recombination. An accompanying volume, Directed Enzyme Evolution: Screening and Selection Methods (ISBN: 1-58829-286-X), is devoted entirely to selection and screening methods that can be applied to the directed evolution of enzymes. Taken together, Directed Evolution Library Creation: Methods and Protocols and Directed Enzyme Evolution: Screening and Selection Methods capture for newcomers and more experienced investigators alike all the key methods for using directed protein evolution to better understand protein structure-function relationships, to discover new enzymes and therapeutic proteins, and to design new assays suitable for specific applications.
700 1 _aArnold, Frances H
_eeditor literario
_4edt
_4http://id.loc.gov/vocabulary/relators/edt
700 1 _aGeorgiou, George
_eeditor literario
_4edt
_4http://id.loc.gov/vocabulary/relators/edt
776 0 8 _iPrinted edition:
_z9781617374715
776 0 8 _iPrinted edition:
_z9781489938534
776 0 8 _iPrinted edition:
_z9781588292858
856 4 0 _uhttps://go.openathens.net/redirector/universidadeuropea.es?url=https://doi.org/10.1385/159259395X
_z(usuarios Universidad Europea de Valencia)
942 _2lcc
_cLE
988 _aSpringer_Protocols_2003
999 _c234921
_d234921