Part:BBa_K2066114:Experience

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Applications of BBa_K2066114

We were able to sequence confirm this part and use it in conjugation with constitutively expressed TetR (K2066022) on psbIA3. We were able to replicate the original synthetic enhancer data when we used two circuits: Bba_K2066118 and Bba_K2066037.

We used a plate reader to find the absolute fluorescent measures over a gradient of induction. We are able to see four discrete steps allowing for control over a multistate output.

Small chemical induction with aTc binds to and inactivates available TetR repressor. When a small amount of aTc is present, the steady state of TetR is still high enough to allow for complete repression of circuit. As you increase the amount of aTc in the circuit, the steady state of activated TetR decreases and the rigidity of the DNA, which previously prevented the looping and activation of transcription, decreases and gives rise to two intermediate steps. At a high aTc concentrations, almost all of the TetR is bound and inactivated and you reach a saturated level of fluorescent expression.

To read further about the synthetic enhancer mechanism, please refer to: http://2016.igem.org/Team:William_and_Mary/Synthetic_Enhancer

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