Team:St Andrews/Demonstrate

Demonstrate

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Demonstration of Function:

We designed this system to detect the effectiveness of novel metabolites as potential lead compounds for antibiotic development. In commercial labs, the way to calibrate the system would require deliberately lysing the cells with a known antibiotic, and then comparing that rate of cell death with the rate of cell death induced by an unknown compound. In early trials of our system, we lysed the cells with a sonicator to ensure that cell death occurred rapidly and completely; however later for later trials we began to use the system in a more realistic sense. We added ampicillin, the mechanism of which interferes with cell wall development and thus induces lysis, to a combined culture of cells from both halves of the system. Then we measured fluorescence over night using a plate reader, the graph of which is below.

The above graph demonstrates that 100 µl of Ampcillin (to a final concentration of 10 mg/mL) lyses the cells which retain the larger component of mNeonGreen (BBa_K2844009) at a rate sufficient for the first 10 strands of the beta barrel to join with the 11th strand (BBa_K2844006).