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Mutagenesis and Functional Selection Protocols for Directed Evolution of Proteins in E. coli
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Evolutionary innovation using EDGE, a system for localized elevated mutagenesis.

Xiao Yi1, Romas Kazlauskas1,2, Michael Travisano1,3

  • 1BioTechnology Institute, University of Minnesota, St. Paul, Minnesota, United States of America.

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|May 1, 2020
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Summary
This summary is machine-generated.

Localized mutagenesis, engineered via Experimental Designed Genic Evolution (EDGE), facilitates rapid adaptation by focusing mutations on specific genes. This approach overcomes the limitations of broad, deleterious mutations for evolutionary innovation.

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Area of Science:

  • Evolutionary biology
  • Genetics
  • Microbiology

Background:

  • Deleterious mutations across the genome can impede evolutionary innovation and adaptation.
  • Existing localized mutagenesis mechanisms (e.g., phase variation) are rare and have limited mutational spectra.

Purpose of the Study:

  • To engineer a platform for studying evolutionary novelty at a single genetic locus.
  • To investigate the potential for adaptation through focused mutagenesis.

Main Methods:

  • Development of the Experimental Designed Genic Evolution (EDGE) platform.
  • Experimental evolution of bacterial resistance to a novel antibiotic using EDGE.

Main Results:

  • Bacterial resistance to a novel antibiotic evolved readily when mutagenesis was focused on a relevant gene using EDGE.
  • A model was proposed explaining the cost-benefit of single-locus adaptation, high mutation rates, limited innovation, and rarity of natural localized mutagenesis.

Conclusions:

  • Localized mutation systems, like EDGE, can drive adaptive evolution efficiently without overly restricting the mutation spectrum.
  • EDGE is a valuable tool for dissecting complex adaptation processes through localized, efficient evolution.