1. Homologous Quorum Sensing Regulatory Circuit: A Dual-Input Genetic Controller for Modulating Quorum Sensing-Mediated Protein Expression in E. coli .
- Author
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Hauk P, Stephens K, Virgile C, VanArsdale E, Pottash AE, Schardt JS, Jay SM, Sintim HO, and Bentley WE
- Subjects
- Acyl-Butyrolactones metabolism, Gene Expression, Gene Expression Regulation, Bacterial, Granulocyte-Macrophage Colony-Stimulating Factor genetics, Granulocyte-Macrophage Colony-Stimulating Factor isolation & purification, Homoserine analogs & derivatives, Homoserine metabolism, Humans, Lactones metabolism, Microorganisms, Genetically-Modified, Plasmids genetics, Recombinant Proteins isolation & purification, Recombinant Proteins metabolism, Escherichia coli genetics, Escherichia coli metabolism, Gene Regulatory Networks, Granulocyte-Macrophage Colony-Stimulating Factor metabolism, Metabolic Engineering methods, Quorum Sensing genetics, Signal Transduction genetics
- Abstract
We developed a hybrid synthetic circuit that co-opts the genetic regulation of the native bacterial quorum sensing autoinducer-2 and imposes an extra external controller for maintaining tightly controlled gene expression. This dual-input genetic controller was mathematically modeled and, by design, can be operated in three modes: a constitutive mode that enables consistent and high levels of expression; a tightly repressed mode in which there is very little background expression; and an inducible mode in which concentrations of two signals (arabinose and autoinducer-2) determine the net amplification of the gene(s)-of-interest. We demonstrate the utility of the circuit for the controlled expression of human granulocyte macrophage colony stimulating factor in an engineered probiotic E. coli . This dual-input genetic controller is the first homologous AI-2 quorum sensing circuit that has the ability to be operated in three different modes. We believe it has the potential for wide-ranging biotechnological applications due its versatile features.
- Published
- 2020
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