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the bacterial uv toggle |
In the beginning, there was a UV Toggle (Collins, 2000). The 2010 MIT iGEM team saw that it was good, and decided to implement the Collins toggle in E.coli to create cells with bistable phenotypes. The team planned for the toggle to control fluorescence and phage polymerization in response to exposing the cells to UV light. After much fine-tuning of the power of the UV exposure, the concentrations of AHL and IPTG, and mask cutting, a pattern of fluorescence finally emerged -- the first image. Then the team decided to make the signal amplify itself, and record a movie to see its propogation. But the team noticed a circle of cell death where the UV exposure had killed some of the cells in the lawn. They decided to make a pLPTa, a low power toggle that still provided bistability, but required less UV power to induce a toggle switch. By site-directed mutagenesis, the team changed the lambda repressor (cI) gene in the Collins pTSMa to a cI that is more sensitive to cleavage by Rec-A, the enzyme activated by UV light exposure. |