Team:Aberdeen Scotland/parameters/invest 4

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University of Aberdeen iGEM 2009

The Input-dependent model

For this model, we no longer assume a trivial process of repression of lacI being liften and QS being turned on. The LacI repression is now a repression / induction hill function with a Kd of 1200 molecules per cell for IPGT forming IPGT lacI complexes. We assume IPGT leaks into the cell from outside. We only set the outside concentration of IPGT. For quorum sensing all we set is an elevated HSL concentration outside which is free to defuse in. We now include a feedback loop into the QS mechanism. To do this we attach a lux box onto the plasmid for producing luxR and luxI, like so:


Input Model Graph 1.jpg

However, we are taking the production strengths of this lux box to be reduced in comparison to our lux box for producing X, Y and cI. The strengths are

Production max for X, Y and cI = 0.44 pops

taken from the iGEM page of ***** and personal communication with **** on ****

Production min for X Y and cI = 0.013 pops

taken from *****

Production max for QS feedback = 0.002 pops

Production min for QS feedback = 0.00015 pops

These values are taken from the paper “ FIOADAOIDFJAOFJASDIOJASOIJADOAJSDAFOJAFOAJSD”

An important new parameter is KP. This is the dissociation constant for “P” activating the lux boxes. P is the luxR/HSL complex . To take a break from the wordy descriptions we will now show some small sections of results for this model, which should be easily interpretable after following the section about the Monte Carlo analysis

Level of KP required for system to function

here we use a Monte Carlo simulation to determine the value of KP in molecules per cell that is required for the system to function (note that the IPTG and HSL input levels added are both 10000 molecules)

Input Model Graph 10.jpg
Input Model Graph 11.jpg

Relationship between KTetR and KCI then KP is 100

We notice that KP must be around 100 molecules to let the system function. We will now explore the relationship between KTetR and KCI at the value of KP=100 molecules per cell. for this simulation we set the HSL and IPTG input levels to 50,000 molecules. In our next section we will try to calculate the minimum levels of HSL and IPTG required to activate the system correctly.