Team:Gaston Day School/Modeling

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Modeling

Our Part: K1749000

A diagram showing the construction of our part. We combined a Cadmium Promoter with a phi-delta activator. We then combined a PO promoter with a GFP reporter. The two parts were combined to create our construct.

Killswitch Model

     We have also begun design work for a passive killswitch. The kill switch will be arabinose-repressible, incorporating a pBAD promoter for a TetR repressor of a TetR repressible promoter of colicin. When the concentration of arabinose drop, the colicin will no longer be repressed and will kill the cell, preventing any contamination.

Construct 1:

Test the function of the arabinose promoter and determine the most effective concentration of arabinose for induction.

Construct 2:

To be used in combination with construct 3 to test the function of the Tet repressor. the complete test plasmid is construct 4.

Construct 3:

To be used in combination with construct 2 to test the function of the Tet repressor. The complete test plasmid is construct 4.

Construct 4:

Test the function of the arabinose inducible promoter linked to the Tet repressor system. Under arabinose induction, GFP production should fall.

Construct 5:

Intermediate construct containing the lethal gene behind the Tet repressible protein.

Construct 6:

The final kill switch design. Under arabinose induction, the clicin will be suppressed using the Tet repressor. If the level of arabinose falls, as would happen in an accidental release, the Tet repression will stop, causing production of colicin and cell death.