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<li class="li2"><a class="navA2" href="https://2019.igem.org/Team:Fudan-TSI/Description">Description</a></li> | <li class="li2"><a class="navA2" href="https://2019.igem.org/Team:Fudan-TSI/Description">Description</a></li> | ||
<li class="li2"><a class="navA2" href="https://2019.igem.org/Team:Fudan-TSI/Design">Design</a></li> | <li class="li2"><a class="navA2" href="https://2019.igem.org/Team:Fudan-TSI/Design">Design</a></li> | ||
− | <li class="li2"><a class="navA2" href="https://2019.igem.org/Team:Fudan-TSI/ | + | <li class="li2"><a class="navA2" href="https://2019.igem.org/Team:Fudan-TSI/Applied_Design" style="white-space:nowrap">Applied Design</a></li> |
− | <li class="li2"><a class="navA2" href="https://2019.igem.org/Team:Fudan-TSI/ | + | <li class="li2"><a class="navA2" href="https://2019.igem.org/Team:Fudan-TSI/Experiments">Experiment</a></li> |
</ul> | </ul> | ||
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<li class="navLi"> | <li class="navLi"> | ||
− | <a class="navA" href="https://2019.igem.org/Team:Fudan-TSI/ | + | <a class="navA" href="https://2019.igem.org/Team:Fudan-TSI/Model">Model</a> |
<div class="n2"> | <div class="n2"> | ||
<ul class="ul2"> | <ul class="ul2"> | ||
<li class="li2"><a class="navA2" href="https://2019.igem.org/Team:Fudan-TSI/Model">Modeling</a></li> | <li class="li2"><a class="navA2" href="https://2019.igem.org/Team:Fudan-TSI/Model">Modeling</a></li> | ||
<li class="li2"><a class="navA2" href="https://2019.igem.org/Team:Fudan-TSI/Software">Software</a></li> | <li class="li2"><a class="navA2" href="https://2019.igem.org/Team:Fudan-TSI/Software">Software</a></li> | ||
+ | <li class="li2"><a class="navA2" href="https://2019.igem.org/Team:Fudan-TSI/Hardware">Hardware</a></li> | ||
</ul> | </ul> | ||
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<ul class="ul2"> | <ul class="ul2"> | ||
<li class="li2"><a class="navA2" style="white-space:nowrap;" href="https://2019.igem.org/Team:Fudan-TSI/Public_Engagement">Education & <br />Public Engagement</a></li> | <li class="li2"><a class="navA2" style="white-space:nowrap;" href="https://2019.igem.org/Team:Fudan-TSI/Public_Engagement">Education & <br />Public Engagement</a></li> | ||
− | <li class="li2"><a class="navA2" style="white-space:nowrap;" href="https://2019.igem.org/Team:Fudan-TSI/ | + | <li class="li2"><a class="navA2" style="white-space:nowrap;" href="https://2019.igem.org/Team:Fudan-TSI/Human_Practices">Integrated <br />Human Practice</a></li> |
<li class="li2"><a class="navA2" href="https://2019.igem.org/Team:Fudan-TSI/Collaborations">Collaboration</a></li> | <li class="li2"><a class="navA2" href="https://2019.igem.org/Team:Fudan-TSI/Collaborations">Collaboration</a></li> | ||
<li class="li2"><a class="navA2" href="https://2019.igem.org/Team:Fudan-TSI/Safety">Safety</a></li> | <li class="li2"><a class="navA2" href="https://2019.igem.org/Team:Fudan-TSI/Safety">Safety</a></li> | ||
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<ul class="ul2"> | <ul class="ul2"> | ||
<li class="li2"><a class="navA2" style="white-space:nowrap;" href="https://2019.igem.org/Team:Fudan-TSI/Team">Team Members</a></li> | <li class="li2"><a class="navA2" style="white-space:nowrap;" href="https://2019.igem.org/Team:Fudan-TSI/Team">Team Members</a></li> | ||
− | <li class="li2"><a class="navA2" href="https://2019.igem.org/Team:Fudan-TSI/Team/ | + | <li class="li2"><a class="navA2" href="https://2019.igem.org/Team:Fudan-TSI/Attributions">Attribution</a></li> |
+ | <li class="li2"><a class="navA2" href="https://2019.igem.org/Team:Fudan-TSI/Acknowledgment">Acknowledgement</a></li> | ||
</ul> | </ul> | ||
</div> | </div> | ||
</li> | </li> | ||
− | <li class="navLi"><a class="navA noSubmenu" href="https://2019.igem.org/Team:Fudan-TSI/ | + | <li class="navLi"><a class="navA noSubmenu" href="https://2019.igem.org/Team:Fudan-TSI/Team/Judging">Judging</a></li> |
</ul> | </ul> | ||
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When we were constructing the plasmid, we encountered a dilemma concerning how RT and Cre should be expressed. Firstly, we thought of putting them both under a same Lac operon so that their expression can be easily induced merely by one kind of inducer—IPTG. Meanwhile, we also considered using different inducers to achieve a more modular design which would be easier to control. As it would take a long time to test which induced expression scheme is better through experiments, we used modelling to test the two constructs. We modelled all the reactions involved and computed the yield of the desired product, i.e. recombined P<sub>target</sub>. Through comparison of the yield acquired using these two induced expression schemes, we decided that the latter scheme should be employed for our system to perform better. <br /><br /> | When we were constructing the plasmid, we encountered a dilemma concerning how RT and Cre should be expressed. Firstly, we thought of putting them both under a same Lac operon so that their expression can be easily induced merely by one kind of inducer—IPTG. Meanwhile, we also considered using different inducers to achieve a more modular design which would be easier to control. As it would take a long time to test which induced expression scheme is better through experiments, we used modelling to test the two constructs. We modelled all the reactions involved and computed the yield of the desired product, i.e. recombined P<sub>target</sub>. Through comparison of the yield acquired using these two induced expression schemes, we decided that the latter scheme should be employed for our system to perform better. <br /><br /> | ||
By common knowledge we can assume that, if the amount of RT and Cre needs to be different to achieve optimal yield, we should choose the second scheme and put them under different operons. On the contrary, if the yield reaches the maximum under the maximum amount of RT and Cre, the first scheme should be chosen. <br /><br /> | By common knowledge we can assume that, if the amount of RT and Cre needs to be different to achieve optimal yield, we should choose the second scheme and put them under different operons. On the contrary, if the yield reaches the maximum under the maximum amount of RT and Cre, the first scheme should be chosen. <br /><br /> | ||
− | In our initial attempt, we found that modelling all the reactions involved is rather difficult, as the reactions are in such a large number and all mixed together. This circumstance makes inspection of the reasonability of our models and parameters impossible. To overcome this issue, we decided to separate these reactions into three minor models and use the steady-state concentration of the substances derived from the previous model as the input of the next model. The three minor models are: <b><i>induced expression model, reverse transcription model and Cre recombination model</i></b>, corresponding to the 3 reaction steps in R-Evolution. The schematic diagram is shown in Fig. 1. | + | In our initial attempt, we found that modelling all the reactions involved is rather difficult, as the reactions are in such a large number and all mixed together. This circumstance makes inspection of the reasonability of our models and parameters impossible. To overcome this issue, we decided to separate these reactions into three minor models and use the steady-state concentration of the substances derived from the previous model as the input of the next model. The three minor models are: <b><i>induced expression model, reverse transcription model and Cre recombination model</i></b>, corresponding to the 3 reaction steps in R-Evolution. The schematic diagram is shown in <a href="#Fig1">Fig. 1</a>. |
</div> | </div> | ||
</div> | </div> |
Revision as of 22:38, 21 October 2019