Project Abstract

Our team engineered a novel genetic toggle switch regulated at the translational level that allowed yeast to express either green or cyan fluorescent protein mutually exclusively. We successfully constructed a fusion of mRNA binding protein and fluorescent protein in yeast characterized using microscopy, a fluorometer and FACS analysis of single cells or whole populations. These results along with parameters from literature were used to predict via deterministic and stochastic models that the probability of successful bistability for our switch was 0.96%, but this could be improved theoretically to a maximum of 51.27% by limiting the range of variation of the most sensitive parameters. The models also implied that co-operativity of binding of the mRNA binding protein to its mRNA stem loop was theoretically essential for generating switch-like behaviour. These results suggest that a genetic toggle switch regulated at the translational level is a viable and novel engineering concept applicable to medicinal, environmental and technological problems.

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