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| <html> | | <html> |
- | <h1 class="title">Deterministic Modeling of a Bacterial Light Recognition Circuit</h1>
| + | {{http://www.brownigem.com/latexconverted.html}} |
- | <h1 id="players"
| + | |
- | >Players</h1
| + | |
- | ><p
| + | |
- | >The following are the players that are used in the modeling of our circuit:</p
| + | |
- | ><h2 id="transcription-factors"
| + | |
- | >Transcription factors</h2
| + | |
- | ><ol
| + | |
- | ><li
| + | |
- | ><p
| + | |
- | >LovTAP</p
| + | |
- | ></li
| + | |
- | ><li
| + | |
- | ><p
| + | |
- | >LovTAP* (After light-induced conformational change)</p
| + | |
- | ></li
| + | |
- | ><li
| + | |
- | ><p
| + | |
- | >tetR</p
| + | |
- | ></li
| + | |
- | ><li
| + | |
- | ><p
| + | |
- | >Mnt</p
| + | |
- | ></li
| + | |
- | ><li
| + | |
- | ><p
| + | |
- | >AraC</p
| + | |
- | ></li
| + | |
- | ><li
| + | |
- | ><p
| + | |
- | >LacI</p
| + | |
- | ></li
| + | |
- | ><li
| + | |
- | ><p
| + | |
- | >CI</p
| + | |
- | ></li
| + | |
- | ><li
| + | |
- | ><p
| + | |
- | >CI434</p
| + | |
- | ></li
| + | |
- | ><li
| + | |
- | ><p
| + | |
- | >GAL4</p
| + | |
- | ></li
| + | |
- | ></ol
| + | |
- | ><h2 id="reporters"
| + | |
- | >Reporters</h2
| + | |
- | ><ol
| + | |
- | ><li
| + | |
- | ><p
| + | |
- | >S1</p
| + | |
- | ></li
| + | |
- | ><li
| + | |
- | ><p
| + | |
- | >S2</p
| + | |
- | ></li
| + | |
- | ><li
| + | |
- | ><p
| + | |
- | >S3</p
| + | |
- | ></li
| + | |
- | ><li
| + | |
- | ><p
| + | |
- | >S4</p
| + | |
- | ></li
| + | |
- | ></ol
| + | |
- | ><h2 id="constants"
| + | |
- | >Constants</h2
| + | |
- | ><p
| + | |
- | >The following are the constants for which we need to find online or determine experimentally:</p
| + | |
- | ><ol
| + | |
- | ><li
| + | |
- | ><p
| + | |
- | ><span class="LaTeX"
| + | |
- | >$\delta = \left\{
| + | |
- | \begin{array}{ll}
| + | |
- | 0 & \mbox{without light}\\
| + | |
- | 1 & \mbox{with light}
| + | |
- | \end{array}
| + | |
- | \right.$</span
| + | |
- | ></p
| + | |
- | ></li
| + | |
- | ><li
| + | |
- | ><p
| + | |
- | ><span class="LaTeX"
| + | |
- | >$c$</span
| + | |
- | > represents the rate of conformation change from LovTAP to LovTAP* under 470nm light.</p
| + | |
- | ></li
| + | |
- | ></ol
| + | |
- | ><h2 id="parameters"
| + | |
- | >Parameters</h2
| + | |
- | ><ol
| + | |
- | ><li
| + | |
- | ><p
| + | |
- | ><span class="LaTeX"
| + | |
- | >$\alpha_x$</span
| + | |
- | > represents the <span class="LaTeX"
| + | |
- | >$x$</span
| + | |
- | >th synthesis rate <span class="LaTeX"
| + | |
- | >$\left(\dfrac{\text{nanomoles}}{\text{min}}\right)$</span
| + | |
- | >.<br
| + | |
- | /> <span class="LaTeX"
| + | |
- | >$\alpha=\left(\text{rate of transcription}\right)\times\left(\text{rate of translation}\right)$</span
| + | |
- | ><br
| + | |
- | /> For promoters for which reliable data is not available, we assume an average <em
| + | |
- | >E. coli</em
| + | |
- | > transcription speed to be 70 nucleotides/second = 4,200 nucleotides/min, and an average translation speed of 40 amino acids/second = 2400, which is then further regulated by the appropriate ribosome binding site, represented as a normalized constant. Thus, we use the following equation to calculate unknown synthesis rates[0]: <span class="LaTeX"
| + | |
- | >$$\alpha = \left(\frac{4200}{\text{gene length}}\right)\times\left(\frac{2400\times\text{RBS Strength}}{\text{protein length}}\right) $$</span
| + | |
- | ></p
| + | |
- | ><p
| + | |
- | ><span class="LaTeX"
| + | |
- | >$\alpha_1$</span
| + | |
- | > LovTAP synthesis rate constant <span class="LaTeX"
| + | |
- | >$= 1.06432\times10^{-13}$</span
| + | |
- | > nanomoles. <span class="LaTeX"
| + | |
- | >$\alpha_2$</span
| + | |
- | > tetR synthesis rate constant <span class="LaTeX"
| + | |
- | >$= 1.15318\times10^{-13}$</span
| + | |
- | > nanomoles. <span class="LaTeX"
| + | |
- | >$\alpha_3$</span
| + | |
- | > Mnt synthesis rate constant <span class="LaTeX"
| + | |
- | >$= 6.0562\times*10^{-13}$</span
| + | |
- | > nanomoles. <span class="LaTeX"
| + | |
- | >$\alpha_4$</span
| + | |
- | > AraC synthesis rate constant <span class="LaTeX"
| + | |
- | >$= 5.99989\times10^{-14}$</span
| + | |
- | > nanomoles. <span class="LaTeX"
| + | |
- | >$\alpha_5$</span
| + | |
- | > LacI synthesis rate constant <span class="LaTeX"
| + | |
- | >$= 3.94791\times10^{-14}$</span
| + | |
- | > nanomoles. <span class="LaTeX"
| + | |
- | >$\alpha_6$</span
| + | |
- | > CI synthesis rate constant <span class="LaTeX"
| + | |
- | >$= 8.93023\times10^{-14}$</span
| + | |
- | > nanomoles. <span class="LaTeX"
| + | |
- | >$\alpha_7$</span
| + | |
- | > CI434 synthesis rate constant <span class="LaTeX"
| + | |
- | >$= 1.2236\times10^{-13}$</span
| + | |
- | > nanomoles. <span class="LaTeX"
| + | |
- | >$\alpha_8$</span
| + | |
- | > [check]SupD synthesis rate constant <span class="LaTeX"
| + | |
- | >$= 1.2236\times10^{-13}$</span
| + | |
- | > nanomoles. <span class="LaTeX"
| + | |
- | >$\alpha_9$</span
| + | |
- | > [check]T7ptag synthesis rate constant <span class="LaTeX"
| + | |
- | >$= 1.2236\times10^{-13}$</span
| + | |
- | > nanomoles. <span class="LaTeX"
| + | |
- | >$\alpha_{10}$</span
| + | |
- | > [check]GAL4 synthesis rate constant <span class="LaTeX"
| + | |
- | >$= 1.06432\times10^{-13}$</span
| + | |
- | > nanomoles.</p
| + | |
- | ><p
| + | |
- | ><span class="LaTeX"
| + | |
- | >$\alpha_{11}$</span
| + | |
- | > S1 synthesis rate constant <span class="LaTeX"
| + | |
- | >$= 9.68992\times10^{-14}$</span
| + | |
- | > nanomoles. <span class="LaTeX"
| + | |
- | >$\alpha_{12}$</span
| + | |
- | > S2 synthesis rate constant <span class="LaTeX"
| + | |
- | >$= 9.68992\times10^{-14}$</span
| + | |
- | > nanomoles. <span class="LaTeX"
| + | |
- | >$\alpha_{13}$</span
| + | |
- | > S3 synthesis rate constant <span class="LaTeX"
| + | |
- | >$= 9.68992\times10^{-14}$</span
| + | |
- | > nanomoles. <span class="LaTeX"
| + | |
- | >$\alpha_{14}$</span
| + | |
- | > S4 synthesis rate constant <span class="LaTeX"
| + | |
- | >$= 9.68992\times10^{-14}$</span
| + | |
- | > nanomoles.</p
| + | |
- | ></li
| + | |
- | ><li
| + | |
- | ><p
| + | |
- | ><span class="LaTeX"
| + | |
- | >$\beta_x $</span
| + | |
- | > represents <span class="LaTeX"
| + | |
- | >$x$</span
| + | |
- | >th basal (un-induced or un-repressed) synthesis rate of a given promoter. We assume that, for all promoters, this is equal to 1% of the synthesis rate constant. <span class="LaTeX"
| + | |
- | >$$\forall x \ \beta_x = 0.1\alpha_x$$</span
| + | |
- | ></p
| + | |
- | ></li
| + | |
- | ><li
| + | |
- | ><p
| + | |
- | ><span class="LaTeX"
| + | |
- | >$\mu_x$</span
| + | |
- | > represents the degradation rate of a given protein. When the degredation rate is unknown, we assume a decay of 0.012 proteins/min (half-life of one hour) <span class="LaTeX"
| + | |
- | >$\mu_1= 0.0453$</span
| + | |
- | > Rate of LovTAP degredation <span class="LaTeX"
| + | |
- | >$(1/sec)$</span
| + | |
- | ><span class="LaTeX"
| + | |
- | >$\mu_2= 0.0453$</span
| + | |
- | > Rate of LovTAP* degredation <span class="LaTeX"
| + | |
- | >$(1/sec)$</span
| + | |
- | ><span class="LaTeX"
| + | |
- | >$\mu_3 = 0.0453$</span
| + | |
- | > tetR degredation <span class="LaTeX"
| + | |
- | >$(1/sec)$</span
| + | |
- | ><span class="LaTeX"
| + | |
- | >$\mu_4= 0.0453$</span
| + | |
- | > Rate of Mnt degredation <span class="LaTeX"
| + | |
- | >$(1/sec)$</span
| + | |
- | ><span class="LaTeX"
| + | |
- | >$\mu_5= 0.0453$</span
| + | |
- | > Rate of AraC degredation <span class="LaTeX"
| + | |
- | >$(1/sec)$</span
| + | |
- | ><span class="LaTeX"
| + | |
- | >$\mu_6= 0.0453$</span
| + | |
- | > Rate of LacI degredation <span class="LaTeX"
| + | |
- | >$(1/sec)$</span
| + | |
- | ><span class="LaTeX"
| + | |
- | >$\mu_7= 0.0453$</span
| + | |
- | > Rate of CI degredation <span class="LaTeX"
| + | |
- | >$(1/sec)$</span
| + | |
- | ><span class="LaTeX"
| + | |
- | >$\mu_8= 0.0453$</span
| + | |
- | > Rate of CI434 degredation <span class="LaTeX"
| + | |
- | >$(1/sec)$</span
| + | |
- | ><span class="LaTeX"
| + | |
- | >$\mu_9= 00453$</span
| + | |
- | > Rate of SupD degredation <span class="LaTeX"
| + | |
- | >$(1/sec)$</span
| + | |
- | ><span class="LaTeX"
| + | |
- | >$\mu_{10}= 0.0453$</span
| + | |
- | > Rate of T7ptag degredation <span class="LaTeX"
| + | |
- | >$(1/sec)$</span
| + | |
- | ><span class="LaTeX"
| + | |
- | >$\mu_{11}= 0.012$</span
| + | |
- | > Rate of T7 polymerase degredation <span class="LaTeX"
| + | |
- | >$(1/sec)$</span
| + | |
- | ><span class="LaTeX"
| + | |
- | >$\mu_{12}= 0.012$</span
| + | |
- | > Rate of GAL4 degredation <span class="LaTeX"
| + | |
- | >$(1/sec)$</span
| + | |
- | ><span class="LaTeX"
| + | |
- | >$\mu_{13}= 0.012$</span
| + | |
- | > Rate of S1 degredation <span class="LaTeX"
| + | |
- | >$(1/sec)$</span
| + | |
- | ><span class="LaTeX"
| + | |
- | >$\mu_{14}= 0.012$</span
| + | |
- | > Rate of S2 degredation <span class="LaTeX"
| + | |
- | >$(1/sec)$</span
| + | |
- | ><span class="LaTeX"
| + | |
- | >$\mu_{15}= 0.012$</span
| + | |
- | > Rate of S3 degredation <span class="LaTeX"
| + | |
- | >$(1/sec)$</span
| + | |
- | ><span class="LaTeX"
| + | |
- | >$\mu_{16}= 0.012$</span
| + | |
- | > Rate of S4 degredation <span class="LaTeX"
| + | |
- | >$(1/sec)$</span
| + | |
- | ></p
| + | |
- | ></li
| + | |
- | ><li
| + | |
- | ><p
| + | |
- | ><span class="LaTeX"
| + | |
- | >$n_x$</span
| + | |
- | > represents the hill coefficient for a given protein. <span class="LaTeX"
| + | |
- | >$n_1 = 1$</span
| + | |
- | > for LovTAP*</p
| + | |
- | ><p
| + | |
- | ><span class="LaTeX"
| + | |
- | >$n_2 = 3$</span
| + | |
- | > for tetR)</p
| + | |
- | ><p
| + | |
- | ><span class="LaTeX"
| + | |
- | >$n_3 = 2$</span
| + | |
- | > for CI</p
| + | |
- | ><p
| + | |
- | ><span class="LaTeX"
| + | |
- | >$n_4 = 2$</span
| + | |
- | > for CI434</p
| + | |
- | ><p
| + | |
- | ><span class="LaTeX"
| + | |
- | >$n_5 = 2$</span
| + | |
- | > Hill coefficient of GAL4</p
| + | |
- | ><p
| + | |
- | ><span class="LaTeX"
| + | |
- | >$n_6 = 2$</span
| + | |
- | > Hill coefficient of AraC</p
| + | |
- | ><p
| + | |
- | ><span class="LaTeX"
| + | |
- | >$n_7 = 2$</span
| + | |
- | > Hill coefficient of T7</p
| + | |
- | ><p
| + | |
- | ><span class="LaTeX"
| + | |
- | >$n_8 = 2$</span
| + | |
- | > Hill coefficient of LacI</p
| + | |
- | ><p
| + | |
- | ><span class="LaTeX"
| + | |
- | >$n_9 = 1$</span
| + | |
- | > Hill coefficient of Mnt</p
| + | |
- | ></li
| + | |
- | ><li
| + | |
- | ><p
| + | |
- | ><span class="LaTeX"
| + | |
- | >$K_x$</span
| + | |
- | > represents, for a given protein, the ligand concentration producing half occupation (ligand concentration occupying half of the binding sites) in nanomoles. This is also the infamous "microscopic dissociation constant". <span class="LaTeX"
| + | |
- | >$K_{d1} = 142$</span
| + | |
- | > for LovTAP*</p
| + | |
- | ><p
| + | |
- | ><span class="LaTeX"
| + | |
- | >$K_{d2} = 0.179$</span
| + | |
- | > for tetR</p
| + | |
- | ><p
| + | |
- | ><span class="LaTeX"
| + | |
- | >$K_{d3} = 50$</span
| + | |
- | > for CI</p
| + | |
- | ><p
| + | |
- | ><span class="LaTeX"
| + | |
- | >$K_{d4} = 40$</span
| + | |
- | > for CI434</p
| + | |
- | ><p
| + | |
- | ><span class="LaTeX"
| + | |
- | >$K_{d5} = 0.5$</span
| + | |
- | > for GAL4</p
| + | |
- | ><p
| + | |
- | ><span class="LaTeX"
| + | |
- | >$K_{d6} = 14$</span
| + | |
- | > for AraC</p
| + | |
- | ><p
| + | |
- | ><span class="LaTeX"
| + | |
- | >$K_{d7} = 2$</span
| + | |
- | > for T7</p
| + | |
- | ><p
| + | |
- | ><span class="LaTeX"
| + | |
- | >$K_{d8} = 800$</span
| + | |
- | >for LacI</p
| + | |
- | ><p
| + | |
- | ><span class="LaTeX"
| + | |
- | >$K_{d9} = 50$</span
| + | |
- | > for Mnt</p
| + | |
- | ></li
| + | |
- | ><li
| + | |
- | ><p
| + | |
- | ><span class="LaTeX"
| + | |
- | >$c$</span
| + | |
- | > <span class="LaTeX"
| + | |
- | >$c_1 = 20.20$</span
| + | |
- | >, which represents the rate of light-induced LovTAP to LovTAP* conformation change <span class="LaTeX"
| + | |
- | >$(1/sec)$</span
| + | |
- | > <span class="LaTeX"
| + | |
- | >$c_2 = 20.20$</span
| + | |
- | > which represents the rate of the reaction SupD + T7ptag <span class="LaTeX"
| + | |
- | >$\rightarrow$</span
| + | |
- | > T7</p
| + | |
- | ></li
| + | |
- | ><li
| + | |
- | ><p
| + | |
- | ><span class="LaTeX"
| + | |
- | >$\delta = \left\{
| + | |
- | \begin{array}{ll}
| + | |
- | 0 & \mbox{without light}\\
| + | |
- | 1 & \mbox{with light}
| + | |
- | \end{array}\right.$</span
| + | |
- | ></p
| + | |
- | ></li
| + | |
- | ></ol
| + | |
- | >
| + | |
| </html> | | </html> |
| | | |
| | | |
- | For a complete list of values and sensitivities, please see our [http://brownigem.com/parameters_2010.ods spreadsheet of parameters and sensitivities]. | + | For a complete list of values and sensitivities, please see our soon-to-be updated [http://brownigem.com/parameters_2010.ods spreadsheet of parameters and sensitivities]. |
As our circuit is relatively complex, we have many factors and thus many parameters with which to describe the function of these factors mathematically.