Abstract
Synthetic biologists aim to design biological systems for a variety of industrial and medical applications, ranging from biofuel to drug production. Synthetic gene circuits regulating efflux pump protein expression can achieve this by driving desired substrates such as biofuels, pharmaceuticals, or other chemicals out of the cell in a precisely controlled manner. However, efflux pumps may introduce implicit negative feedback by pumping out intracellular inducer molecules that control gene circuits, which then can alter gene circuit function. Therefore, synthetic gene circuits must be carefully designed and constructed for precise efflux control. Here, we provide protocols for quantitatively modeling and building synthetic gene constructs for efflux pump regulation.
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Acknowledgments
The authors would like to thank W.-K. Huh for kindly sharing the PDR5::GFP fusion, and M. Bennett, M. Lorenz, G. May, T. F. Cooper, G. Peng for helpful discussions, and Matthew Wu for editing the modeling section of the chapter. This research was supported by the NIH Director’s New Innovator Award (1DP2 OD006481− 01) and an NIGMS Maximizing Investigators’ Research Award (MIRA, 1R35GM122561) to G.B.; by an NSERC Postdoctoral Fellowship [Grant no: PDF-453977−2014] to D.C.; by the University of Texas Graduate School of Biomedical Sciences at Houston to J.D.; by Program # 1326 of the Ministry of Education and Science, Russian Federation to D.N.; and by the Laufer Center for Physical & Quantitative Biology. Daniel A. Charlebois and Junchen Diao contributed equally to this work.
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Charlebois, D.A., Diao, J., Nevozhay, D., Balázsi, G. (2018). Negative Regulation Gene Circuits for Efflux Pump Control. In: Braman, J. (eds) Synthetic Biology. Methods in Molecular Biology, vol 1772. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7795-6_2
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DOI: https://doi.org/10.1007/978-1-4939-7795-6_2
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