Team:WITS-CSIR SA

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<p>  Bacterial chemotaxis is controlled via a signalling cascade, where CheZ is a protein integral in the directed movement of bacteria towards a stimulus. The aim was to control chemotaxis such that bacteria will be attracted to a defined substance followed by the ability to travel back to another stimulus at the start location, upon activation of an IPTG inducible toggle switch. Two riboswitches were used to control the translation of a CheZ fluorescent protein fusion, the first sensitive to theophylline and the second to atrazine. Fluorometry was used to prove the activation of the riboswitches. A theophylline concentration of 1.5mM resulted in the highest expression of the fusion protein. Motility experiments indicated that CheZ mutants regained motility in the presence of theophylline. Since riboswitches can be engineered for many substances, this system has possible applications as a networking template in multiple situations, be they industrial or medical.</p>
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Revision as of 19:53, 3 September 2011

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Bacterial chemotaxis is controlled via a signalling cascade, where CheZ is a protein integral in the directed movement of bacteria towards a stimulus. The aim was to control chemotaxis such that bacteria will be attracted to a defined substance followed by the ability to travel back to another stimulus at the start location, upon activation of an IPTG inducible toggle switch. Two riboswitches were used to control the translation of a CheZ fluorescent protein fusion, the first sensitive to theophylline and the second to atrazine. Fluorometry was used to prove the activation of the riboswitches. A theophylline concentration of 1.5mM resulted in the highest expression of the fusion protein. Motility experiments indicated that CheZ mutants regained motility in the presence of theophylline. Since riboswitches can be engineered for many substances, this system has possible applications as a networking template in multiple situations, be they industrial or medical.

  1. Concept

    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.

  2. Meet the Team

  3. Slide 6

  4. Slide 6

  5. Slide 6