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Genetic selection for small molecule production in competitive microfluidic droplets

View ORCID ProfileLarry J. Millet, View ORCID ProfileJessica M. Velez, Joshua K. Michener
doi: https://doi.org/10.1101/469007
Larry J. Millet
Biosciences Division, Oak Ridge National Laboratory, Oak Ridge, TN, 37830The Joint Research Activity Unit of The Bredesen Center for Interdisciplinary Research and Graduate Education, University of Tennessee, Knoxville, TN, 37996
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  • ORCID record for Larry J. Millet
Jessica M. Velez
Biosciences Division, Oak Ridge National Laboratory, Oak Ridge, TN, 37830
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Joshua K. Michener
Biosciences Division, Oak Ridge National Laboratory, Oak Ridge, TN, 37830
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  • For correspondence: michenerjk@ornl.gov
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Abstract

Biosensors can be used to screen or select for small molecule production in engineered microbes. However, mutations to the biosensor that interfere with accurate signal transduction are common, producing an excess of false positives. Strategies have been developed to avoid this limitation by physically separating the production pathway and biosensor, but these approaches have only been applied to screens, not selections. We have developed a novel biosensor-mediated selection strategy using competition between co-cultured bacteria. When applied to biosynthesis of cis,cis-muconate, we show that this strategy yields a selective advantage to producer strains that outweighs the costs of production. By encapsulating the competitive co-cultures into microfluidic droplets, we successfully enriched for muconate-producing strains from a large population of control non-producers. Facile selections for small molecule production will increase testing throughput for engineered microbes and allow for the rapid optimization of novel metabolic pathways.

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted November 29, 2018.
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Genetic selection for small molecule production in competitive microfluidic droplets
Larry J. Millet, Jessica M. Velez, Joshua K. Michener
bioRxiv 469007; doi: https://doi.org/10.1101/469007
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Genetic selection for small molecule production in competitive microfluidic droplets
Larry J. Millet, Jessica M. Velez, Joshua K. Michener
bioRxiv 469007; doi: https://doi.org/10.1101/469007

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