Model

The whole cell model is the combination of the Quorum sensing, Integrase and XOR modules. The model shows the behaviour of the a single cell in response to incoming signals. The model enables us to understand the effect of leakiness, cross-talk and their combinations on the whole system.

Chemical Species

Reactions

For the Lux system

$$ \begin{align} &\rightarrow LuxR \\ Lux-AHL+LuxR & \leftrightarrow RLux\\ RLux+RLux &\leftrightarrow DRLux\\ DRLux+P_{luxOFF} & \leftrightarrow P_{luxON}\\ P_{luxON}&\rightarrow P_{luxON}+mRNA_{Bxb1}\\ mRNA_{Bxb1}&\rightarrow Bxb1\\ AHL &\rightarrow \\ LuxR &\rightarrow \\ RLux &\rightarrow\\ DRLux &\rightarrow\\ mRNA_{Bxb1} &\rightarrow\\ Bxb1 &\rightarrow \end{align}$$

For the Las system

\begin{align} &\rightarrow LasR \\ Las-AHL+LasR & \leftrightarrow RLas \\ RLas+RLas & \leftrightarrow DRLas\\ DRLas+P_{LasOFF} & \leftrightarrow P_{LasON}\\ P_{LasON}&\rightarrow P_{LasON}+mRNA_{\phi C31}\\ Las-AHL &\rightarrow \\ LasR &\rightarrow \\ RLas &\rightarrow\\ DRLas &\rightarrow\\ mRNA_{\phi C31} &\rightarrow \\ \phi C31 &\rightarrow \\ \end{align}

Applying mass action kinetic laws, we obtain the following set of differential equations. $$\begin{align*} \frac{d[Lux-AHL]}{dt} &= k_{-RLux}[R_{Lux}]-k_{RLux}[Lux-AHL][LuxR]-d_{Lux-AHL}[Lux-AHL]\\ \frac{d[LuxR]}{dt} &= \alpha_{LuxR} -k_{RLux}[Lux-AHL][LuxR] + k_{-RLux}[RLux] - d_{LuxR}[LuxR] \\ \frac{d[RLux]}{dt} &= k_{RLux}[Lux-AHL][LuxR] - k_{-RLux}[RLux] - 2 k_{DRLux} [RLux]^2 + 2 k_{-DRLux} [DRLux] - d_{RLux} [RLux] \\ \frac{d[DRLux]}{dt} &= k_{DRLux} [RLux]^2 - k_{-DRLux} [DRLux] - d_{DRLux} [DRLux] \\ \frac{d[P_{LuxON}]}{dt} &= k_{P_{LuxON}} [P_{LuxOFF}][DRLux] - k_{-P_{LuxON}} [P_{LuxON}]\\ \frac{d[mRNA_{Bxb1}]}{dt} &= L_{P_{Lux}} + k_{mRNA_{Bxb1}} [P_{LuxON}] - d_{mRNA_{Bxb1}} [mRNA_{Bxb1}]\\ \frac{d[Bxb1]}{dt} &= k_{mRNA_{Bxb1}} [mRNA_{Bxb1}] - d_{Bxb1}[Bxb1]\\ \end{align*}$$

The same holds true for the Las system.

Characterization: K_mLux and K_mLas

Data

For the Quorum sensing module we used established experimentally determined parameters for the rate of formation of RLux (reference). Since, in the literature the other parameters were estimated or fitted to their data, we decided to determine the parameters specific to our system. Hence, we used our data for the remaining parameters. Our data was mainly a transfer function of normalized GFP concentration as a function of input Lux-AHL concentrations. (link to data)

Assumptions

From the original set of reactions, we reduce the rate of production of mRNA_Bxb1 as a Hill function of RLux instead of Mass action kinetics in terms of P_LuxON and P_LuxOFF.

Assumption A

We assumed that the dimerization of RLux to DRLux is quick. Quasi steady state approximation (QSSA) as follows

$$\frac{d[DRLux]}{dt} = k_{DRLux} [RLux]^2 - k_{-DRLux} [DRLux] - d_{DRLux} [DRLux] \approx 0\\$$

Assumption B

Further, from literature, we found that DRLux is specific to DNA and the dissociation constant is low (k_m = 0.1nM) {Reference}. Therefore, we using QSSA again,

$$\frac{d[P_{LuxON}]}{dt} = k_{P_{LuxON}} [P_{LuxOFF}][DRLux] - k_{-P_{LuxON}} [P_{LuxON}] \approx 0\\$$

Solving, we get the rate of production of mRNA_Bxb1 as $$\frac{d[mRNA_{Bxb1}]}{dt} = L_{P_{Lux}} + \frac{K_{mRNA_{Bxb1}}RLux^2}{K_{mLux} + RLux^2 }- d_{mRNA_{Bxb1}} [mRNA_{Bxb1}]\\$$

where

$$K_{mLux} = \frac {k_{-P_{LuxON}}}{k_{P_{LuxON}}}.\frac {k_{-DRLux} + d_{DRLux}}{k_{DRLux}}$$

is a lumped parameter which we fitted to our data.

Similarly, lumped parameter K_mLas was derived for the las system and fitted to a transfer function of normalized GFP concentration as a function of input Las-AHL.

Parameter fitting

We used MEIGO Toolbox to fit the parameters to the transfer functions.

Range of validity of the assumptions

These assumptions hold true for all input Lux-AHL and Las-AHL concentrations.