Phenotypic diversity as a tool to guide and optimize random strain improvement approaches

[摘要] (cont.) We modified our divergence quantification protocol to test whether individual amino acids in the alpha subunit could be experimentally considered as determinants for diversity. We showed that not only can single residues be probed individually, but also that, by testing the phenotypic diversity produced by saturation mutagenesis at different positions, we could find regions and functionalities that are promising for further studies. We proposed this as a novel way for reducing the search space in a particular target for the purpose of increasing the quality of a library. What started as an effort to improve upon transcriptional engineering, soon evolved into a general approach to optimize random search-based methods for isolating traits of interest. We demonstrated the use of this approach for guiding the construction of transcriptional engineering libraries, and in addition outlined the conceptual framework for extending this work to any genetic library. As such, the work of this thesis served both theoretical and practical goals, and furthered the understanding of how evolution can be exploited in the laboratory

[发布机构] Massachusetts Institute of Technology