Construction of a Genetic Multiplexer to Toggle between Chemosensory Pathways in Escherichia coli

Many applications require cells to switch between discrete phenotypic states. Here, we harness the FimBE inversion switch to flip a promoter, allowing expression to be toggled between two genes oriented in opposite directions. The response characteristics of the switch are characterized using two-color cytometry. This switch is used to toggle between orthogonal chemosensory pathways by controlling the expression of CheW and CheW*, which interact with the Tar (aspartate) and Tsr* (serine) chemoreceptors, respectively. CheW* and Tsr* each contain a mutation at their protein–protein interface such that they interact with each other. The complete genetic program containing an arabinose-inducible FimE controlling CheW/CheW* (and constitutively expressed tar/tsr*) is transformed into an Escherichia coli strain lacking all native chemoreceptors. This program enables bacteria to swim toward serine or aspartate in the absence or in the presence of arabinose, respectively. Thus, the program functions as a multiplexer with arabinose as the selector. This demonstrates the ability of synthetic genetic circuits to connect to a natural signaling network to switch between phenotypes.

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► An invertase (FimE) is used to switch between two chemotaxis phenotypes.
► Stochastic and incomplete switching is observed by two-color flow cytometry.
► A genetic program is constructed, where an arabinose-inducible system switches between two orthogonal CheW proteins, which interact with different tar-like chemoreceptors, to change the chemical into which the bacteria swim.