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Team:SDU-Denmark/Tour40
From 2014.igem.org
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<h4>Controlling the Tet promoter</h4> | <h4>Controlling the Tet promoter</h4> | ||
| - | <a class="popupImg alignRight" style="width:200px" target="_blank" href="https://static.igem.org/mediawiki/2014/b/b0/2014SDUresults3.png" title="Plate containing 0 ng/mL doxycycline plated with <i>E. coli</i> WT and <i>E. coli</i> expressing GFP controlled by a constitutive promoter, pTet-TeR(+LVA) and pTet-TetR(no LVA)."> | + | <a class="popupImg alignRight" style="width:200px" target="_blank" href="https://static.igem.org/mediawiki/2014/b/b0/2014SDUresults3.png" title="Figure 1: Plate containing 0 ng/mL doxycycline plated with <i>E. coli</i> WT and <i>E. coli</i> expressing GFP controlled by a constitutive promoter, pTet-TeR(+LVA) and pTet-TetR(no LVA)."> |
<img src="https://static.igem.org/mediawiki/2014/e/e6/2014SDUresults4.png" style="width:200px" /> | <img src="https://static.igem.org/mediawiki/2014/e/e6/2014SDUresults4.png" style="width:200px" /> | ||
| - | Plate containing 0 ng/mL doxycycline plated with <i>E. coli</i> WT and <i>E. coli</i> expressing GFP controlled by a constitutive promoter, pTet-TeR(+LVA) and pTet-TetR(no LVA). | + | Figure 1: Plate containing 0 ng/mL doxycycline plated with <i>E. coli</i> WT and <i>E. coli</i> expressing GFP controlled by a constitutive promoter, pTet-TeR(+LVA) and pTet-TetR(no LVA). |
</a> | </a> | ||
| - | <a class="popupImg alignRight" style="width:200px" target="_blank" href="https://static.igem.org/mediawiki/2014/4/4f/2014SDUresults5.png" title="Plate containing 200 ng/mL doxycycline plated with <i>E. coli</i> WT and <i>E. coli</i> expressing GFP controlled by a constitutive promoter, pTet-TeR(+LVA) and pTet-TetR(no LVA)."> | + | <a class="popupImg alignRight" style="width:200px" target="_blank" href="https://static.igem.org/mediawiki/2014/4/4f/2014SDUresults5.png" title="Figure 2: Plate containing 200 ng/mL doxycycline plated with <i>E. coli</i> WT and <i>E. coli</i> expressing GFP controlled by a constitutive promoter, pTet-TeR(+LVA) and pTet-TetR(no LVA)."> |
<img src="https://static.igem.org/mediawiki/2014/1/19/2014SDUresults6.png" style="width:200px" /> | <img src="https://static.igem.org/mediawiki/2014/1/19/2014SDUresults6.png" style="width:200px" /> | ||
| - | Plate containing 200 ng/mL doxycycline plated with <i>E. coli</i> WT and <i>E. coli</i> expressing GFP controlled by a constitutive promoter, pTet-TeR(+LVA) and pTet-TetR(no LVA). | + | Figure 2: Plate containing 200 ng/mL doxycycline plated with <i>E. coli</i> WT and <i>E. coli</i> expressing GFP controlled by a constitutive promoter, pTet-TeR(+LVA) and pTet-TetR(no LVA). |
</a> | </a> | ||
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<br><br><br> | <br><br><br> | ||
<div> | <div> | ||
| - | <a class="popupImg alignCenter" style="width:500px" target="_blank" href="https://static.igem.org/mediawiki/2014/1/12/2014SDUresults2.png" title="Dose response to doxycycline."> | + | <a class="popupImg alignCenter" style="width:500px" target="_blank" href="https://static.igem.org/mediawiki/2014/1/12/2014SDUresults2.png" title="Figure 3: Dose response to doxycycline."> |
<img src="https://static.igem.org/mediawiki/2014/1/12/2014SDUresults2.png" style="width:500px" /> | <img src="https://static.igem.org/mediawiki/2014/1/12/2014SDUresults2.png" style="width:500px" /> | ||
| - | Dose response to doxycycline. | + | Figure 3: Dose response to doxycycline. |
</a> | </a> | ||
</div> | </div> | ||
Revision as of 04:53, 17 October 2014
Results
"I know that I am intelligent, because I know that I know nothing." – Socrates
The next few pages will guide you through the results of the characterization of our submitted parts. On this page, you will find short description of our results, leading you to the final result. We hope you will dig deeper into the results of our systems. After the need to prioritize subprojects, the aim of our project was to get E. coli to express a self-designed nutritional protein tasting of lemon, controlled by an inducible promoter.
Synthesis of self-designed protein OneProt
Pictures of worms/WB
Growth curve
Controlling the Tet promoter
Figure 1: Plate containing 0 ng/mL doxycycline plated with E. coli WT and E. coli expressing GFP controlled by a constitutive promoter, pTet-TeR(+LVA) and pTet-TetR(no LVA).
Figure 2: Plate containing 200 ng/mL doxycycline plated with E. coli WT and E. coli expressing GFP controlled by a constitutive promoter, pTet-TeR(+LVA) and pTet-TetR(no LVA).
In order for us to be able to control the amount of OneProt produced, we need a promoter that can be controlled. For this, we put OneProt under expressional control by pTet. Besides investigating the controllable expression, we also investigated the influence of the LVA tag on TetR. We proved that the inhibition of pTet by TetR is correlated with the concentration of inducer – the more inducer in the less inhibition of pTet. Comparing the expression by pTet controlled by TetR with and without LVA tag shows that there is a higher expression upon induction, using TetR with LVA than using TetR without LVA. We also show that the pTet is leaky.
Figure 3: Dose response to doxycycline.
Flavor improvement
The thought of eating E. coli does not sound that delicious – and that is why we want our OneProt to taste like lemon. The cloning did not succeed, however, we have been able to compare an odor free E. coli strain with an indole producing E. coli strain, which is the compound that gives E. coli in LB media its characteristic odor. We have proved that
Added parts and devices
To the great iGEM registry of biobricks we have added 3 basic parts, 2 regulatory devices and 4 regulable production devices.
