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Team:BostonU/QS
From 2013.igem.org
Quorum Sensing
Quorum sensing is a system that controls population density using stimulus and response. These types of systems can be commandeered for use in synthetic biology and re-engineering to control gene expression in genetic circuits that are implanted in bacteria. The bacteria utilize quorum sensing to coordinate gene expression. To do so, the bacteria produce and secrete signaling molecules. These bacteria have receptors that detect the signaling molecule. When the signaling molecule binds to the receptor, it induces gene expression. The presence of one or more proteins will induce a promoter that controls the production of signal proteins, such as fluorescent proteins, or of other functional genes to link synthetic genetic circuits together.
A LuxIR-type quorum sensing system has been detected in Chromobacterium violaceum (Stauff et al., 2011). The goal is to introduce a new LuxR/I-like quorum sensing system to synthetic biology via MoClo. Chromobacterium violaceum, a gram-negative bacteria found in flora from water and soil in tropical and subtropical regions, uses the CviR/I system. The CviR/I system is homologous to the LuxR/I system. This can be done by cloning and characterizing CviR/I and pVioA, a promoter with CviR binding site) into E.coli.
pVioA as a Constitutive Promoter
The graphs below show MEFL converted RFP measurements (done using the TASBE tools). The data is binned by fluorescence intensity and each bin also includes the cell count. The bottom graph shows the same data as the top one above but it is ordered by cell count, from highest (dark purple) to lowest (pale purple).
We can now see that within the population of cells, the smallest binned group shows the highest level of expression. This may be due to higher copy counts of plasmids within that subpopulation of cells.
