Abstract
SCHEMA is a method of designing protein recombination libraries that contain a large fraction of functional proteins with a high degree of mutational diversity. In the previous chapter, we illustrated the method for designing libraries by swapping contiguous sequence elements. Here, we introduce the NCR (“noncontiguous recombination”) algorithm to identify optimal designs for swapping elements that are contiguous in the 3-D structure but not necessarily in the primary sequence. To exemplify the method, NCR is used to recombine three fungal cellobiohydrolases (CBH1s) to produce a library containing more than 500,000 novel chimeric sequences.
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Acknowledgments
The authors acknowledge funding from the Institute for Collaborative Biotechnologies through grant W911NF-09-D-0001 from the US Army Research Office and the National Central University, Taiwan, through a Cooperative Agreement for Energy Research Collaboration. M.A.S. is supported by a Resnick Sustainability Institute fellowship.
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Smith, M.A., Arnold, F.H. (2014). Noncontiguous SCHEMA Protein Recombination. In: Gillam, E., Copp, J., Ackerley, D. (eds) Directed Evolution Library Creation. Methods in Molecular Biology, vol 1179. Springer, New York, NY. https://doi.org/10.1007/978-1-4939-1053-3_23
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DOI: https://doi.org/10.1007/978-1-4939-1053-3_23
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