Team:KULeuven/Project/Inverter

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Pictogram inverter.png

Contents

Invertimer

BioBricks

Project inverter.jpg

Components

The input signal for the inverter is filtered by using the T7 RNA polymerase promoter (BBa_I712074) and RiboLock3d (BBa_J23078). The inverter's output signal is the LuxI protein, encoded by BBa_C0061. The basis of the inverter system is the LacI repressor molecule, encoded by BBa_C0012, and the lac promotor (BBa_R0011).

Action

When there is a consistent input signal that can penetrate the filter, LacI is produced from the T7 promoter with the adjacent RiboLock. This LacI (BBa_C0012) contains a C-terminal LVA tag for faster degradation and thus better responsiveness. LacI inhibits transcription from the lac promoter just downstream, acting as an inverter for the output LuxI signal. When no (or not enough, because of the filter) signal is present, the LacI repressor will not be made and LuxI will be produced. It is thus clear that when the filter system produces an input signal for the inverter, there will be repression of LuxI production. Otherwise, when no input signal from the filter system is present, LuxI will be produced.

3OC6HSL.gif

When LuxI is present, it functions as a timer, slowly producing the quorum-sensing molecule 3OC6HSL from hexanoyl-ACP and SAM. 3OC6HSL has a very long lifetime if no quorum-quenching lactonase is present and will accumulate. Not only inside the cell but also in the medium.

3OC6HSL is the link to the next device in our system, CellDeath