Abstract
Utilization of structural information from homologous proteins to design novel enzymes is one of the practical applications of structural biology. Structure-based protein engineering is a more reasonable strategy compared with general random mutagenesis. Here, we describe a useful method for production of a series of mutant enzymes based on a cell-free translation system. We employed PCR-mediated in vitro site-directed mutagenesis in combination with wheat-embryo cell-free protein synthesis to establish a high-throughput system. The efficient generation of a series of mutant enzymes facilitates high-throughput screening of functionally improved enzymes.
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Kanno, T., Tozawa, Y. (2010). Protein Engineering Accelerated by Cell-Free Technology. In: Endo, Y., Takai, K., Ueda, T. (eds) Cell-Free Protein Production. Methods in Molecular Biology, vol 607. Humana Press. https://doi.org/10.1007/978-1-60327-331-2_9
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DOI: https://doi.org/10.1007/978-1-60327-331-2_9
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Print ISBN: 978-1-60327-330-5
Online ISBN: 978-1-60327-331-2
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