This project on fluorescence complementation involves the joining of two proteins, Jun and Fos, each fused to a half terminus of YFP. Both of these chimeric proteins were fused to a beta gene that codes for a membrane protein. Then, two cell populations - one expressing Jun-beta-YFPN and the other Fos-beta-YFPC - were combined, ideally resulting in the fusion of the Jun and Fos proteins on the cell membrane. The two halves of the YFP protein would bind as well, giving rise to fluorescence.
Background
Methods and Materials
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Our project concerns the development of a repressilator coupled to quorum sensing as described by Jordi Garcia-Ojalvo, Michael B. Elowitz and Steven H. Strogatz in PNAS. The Elowitz Repressilator attempts to decrease the loss of standard oscillations that previous repressilators faced by utilizing quorum sensing as a means of synchronizing and maintaining standard oscillations. We expanded on this theory by adding YFP and CFP to allow a visual confirmation of the oscillation, and a TetR promoter in front of the LuxR gene and cI after the LuxI gene. Our hopes were that this would assist in standardizing the oscillation of the bacteria.
Theory Behind the Oscillator
Methods and Materials
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