<p>The first construct, called Motility factor – Send, will allow the bacterium to chemotactically search the area of the petri-dish for tetracycline. Upon finding the source of the tetracycline, the bacteria will express a reporter protein.</p>
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<p>The first construct, called Motility factor – Send, will allow the bacterium to chemotactically search the area of the petri-dish for tetracycline. Upon finding the source of the tetracycline, the bacteria will express a reporter protein.Once the source of the tetracycline has been found by the bacterium, a tetracycline-sensitive Toggle Switch construct will be activated. This will turn off the first construct, and turn on the third.The third construct is called Motility factor – Return. The activation of this construct and the deactivation of the Motility factor - Send will alter the chemotactic behaviour of the bacterium and make the bacterium chemotactically responsive to theophylline, instead of tetracycline. The source of theophylline will be located at the initial point of departure. At this location, the cells that have returned can be detected via a second reporter protein. The presence of this protein will be indicative of the presence of tetracycline in the area.</p>
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<p>Once the source of the tetracycline has been found by the bacterium, a tetracycline-sensitive Toggle Switch construct will be activated. This will turn off the first construct, and turn on the third.</p>
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<p>The third construct is called Motility factor – Return. The activation of this construct and the deactivation of the Motility factor - Send will alter the chemotactic behaviour of the bacterium and make the bacterium chemotactically responsive to theophylline, instead of tetracycline. The source of theophylline will be located at the initial point of departure. At this location, the cells that have returned can be detected via a second reporter protein. The presence of this protein will be indicative of the presence of tetracycline in the area.</p>
As a group of students from Wits University in Johannesburg, South Africa, we have set out to genetically reprogram the chemotactic behaviour of E. coli, through the use of synthetic riboswitches. This will enable bacteria to search a defined area for a particular ligand and return to a set location (the starting point), where they can report on their findings. As a proof of principle, we want to test the ability of the reprogrammed cells to locate atrazine/theophylline on a petri-dish, and return to the initial point of departure for reporting. To do this we have three constructs that will make the bacteria toggle between two states of chemotactic responsiveness to different substances.
The first construct, called Motility factor – Send, will allow the bacterium to chemotactically search the area of the petri-dish for tetracycline. Upon finding the source of the tetracycline, the bacteria will express a reporter protein.Once the source of the tetracycline has been found by the bacterium, a tetracycline-sensitive Toggle Switch construct will be activated. This will turn off the first construct, and turn on the third.The third construct is called Motility factor – Return. The activation of this construct and the deactivation of the Motility factor - Send will alter the chemotactic behaviour of the bacterium and make the bacterium chemotactically responsive to theophylline, instead of tetracycline. The source of theophylline will be located at the initial point of departure. At this location, the cells that have returned can be detected via a second reporter protein. The presence of this protein will be indicative of the presence of tetracycline in the area.