505-00 Library generation and auxotrophic selection assays in Escherichia coli and Thermus thermophilus / Jorg Claren, Thomas Schwab, and Reinhard Sterner. Rational and semirational protein design / Ivan V. Korendovych -- Computational analysis of protein tunnels and channels / Jan Brezovsky, Barbora Kozlikova, and Jiri Damborsky -- YASARA : a tool to obtain structural guidance in biocatalytic investigations / Henrik Land and Maria Svedendahl Humble -- Computational library design protocol for rapid improvement of protein stability : FRESCO / Hein J. Wijma, Maximilian J. L. J. Furst, and Dick B. Janssen -- Directed evolution of proteins based on mutational scanning / Carlos G. Acevedo-Rocha, Matteo Ferla, and Manfred T. Reetz -- Brief guide to the high-throughput expression of directed evolution libraries / Ana Luısa Ribeiro, Mario Mencıa, and Aurelio Hidalgo -- Library growth and protein expression : optimal and reproducible microtiter plate expression of recombinant enzymes in E. coli using MTP shakers / Sandy Schmidt, Mark Dorr, and Uwe T. Bornscheuer -- Normalized screening of protein engineering libraries by split-GFP crude cell extract quantification / Javier Santos-Aberturas, Mark Dorr, and Uwe T. Bornscheuer -- Functional analysis of membrane proteins produced by cell-free translation / Srujan Kumar Dondapati, Doreen A. Wustenhagen, and Stefan Kubick -- Practical considerations regarding the choice of the best high-throughput assay / Carolin Mugge and Robert Kourist -- High-throughput screening assays for lipolytic enzymes / Alexander Fulton, Marc R. Hayes, Ulrich Schwaneberg, Jorg Pietruszka, and Karl-Erich Jaeger -- Continuous high-throughput colorimetric assays for α-transaminases / Egon Heuson, Jean-Louis Petit, Franck Charmantray, Veronique de Berardinis, and Thierry Gefflaut -- Colorimetric high-throughput screening assays for the directed evolution of fungal laccases / Isabel Pardo and Susana Camarero -- Directed coevolution of two cellulosic enzymes in Escherichia coli based on their synergistic reactions / Min Liu, Lidan Ye, and Hongwei Yu -- Program-guided design of high-throughput enzyme screening experiments and automated data analysis/evaluation / Mark Dorr and Uwe T. Bornscheuer -- Solid-phase agar plate assay for screening amine transaminases / Martin S. Weiss, Uwe T. Bornscheuer, and Matthias Hohne -- Ultrahigh-throughput screening of single-cell lysates for directed evolution and functional metagenomics / Fabrice Gielen, Pierre-Yves Colin, Philip Mair, and Florian Hollfelder -- Isolation of pH-sensitive antibody fragments by fluorescence-activated cell sorting and yeast surface display / Christian Schroter, Simon Krah, Jan Beck, Doreen Konning, Julius Grzeschik, Bernhard Valldorf, Stefan Zielonka, and Harald Kolmar -- Library generation and auxotrophic selection assays in Escherichia coli and Thermus thermophilus / Jorg Claren, Thomas Schwab, and Reinhard Sterner.
Summary:
This volume details basic and advanced protocols for both stages of protein engineering: the library design phase and the identification of improved variants by screening and selection. Chapters focus on enzyme engineering using rational and semi-rational approaches. Written in the highly 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 laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, Protein Engineering: Methods and Protocols aims to aid scientists in the planning and performance of their experiments.
This resource is supported by the Institute of Museum and Library Services under the provisions of the Library Services and Technology Act as administered by State Library of Iowa.