Team:KU Leuven/Project/HoneydewSystem

From 2013.igem.org

(Difference between revisions)
Line 127: Line 127:
     <div class="row-fluid">
     <div class="row-fluid">
     <div class="span12">
     <div class="span12">
-
       <p align="justify">Tekst over flux balance analysis</p>
+
       <p align="justify">We ran the Flux Balancing Analysis using the COBRA Toolbox for MATLAB. With this analysis we checked whether the growth rate of our cells will be affected when we introduce our methyl salicylate system.</p>
     </div>
     </div>
     </div>
     </div>

Revision as of 21:28, 4 October 2013

iGem

Secret garden

Congratulations! You've found our secret garden! Follow the instructions below and win a great prize at the World jamboree!


  • A video shows that two of our team members are having great fun at our favourite company. Do you know the name of the second member that appears in the video?
  • For one of our models we had to do very extensive computations. To prevent our own computers from overheating and to keep the temperature in our iGEM room at a normal level, we used a supercomputer. Which centre maintains this supercomputer? (Dutch abbreviation)
  • We organised a symposium with a debate, some seminars and 2 iGEM project presentations. An iGEM team came all the way from the Netherlands to present their project. What is the name of their city?

Now put all of these in this URL:https://2013.igem.org/Team:KU_Leuven/(firstname)(abbreviation)(city), (loose the brackets and put everything in lowercase) and follow the very last instruction to get your special jamboree prize!

tree ladybugcartoon

We aim to achieve a sustainable way to reduce the damage caused by aphid pests, and offer an effective alternative for insecticides. Our modified E. coli, the BanAphid (meaning ‘to ban aphids’ as well as with ‘benefits’), would imitate insecticides by using the aphid’s own alarm pheromone, E-β-farnesene, (EBF) to repel them off the plant. On top of that we want to attract aphid predators such as the ladybug by using methyl salicylate (MeS), a phytohormone. This way we make sure the aphids are thoroughly removed from the plant.
We have established what might be possible hurdles in introducing this system in the agricultural industry. First we have to make sure that the plant cell’s metabolism is not over burdened. Besides that we have to take into account that aphids might habituate to constitutive expression of EBF (De Vos et al., 2010, Kunert et al., 2010). Finally, we do not want to attract the aphid’s natural predators when they are not needed.

E-β-Farnesene

EBF serves as the most universal aphid alarm pheromone. It is released from the cornicles of the aphids to warn others against upcoming danger, such as the natural predators of aphids. Because of the fact that EBF is highly susceptible to oxidation, we want to make our BanAphids produce EBF regularly.

Methyl Salicylate - Wetlab

Methyl salicylate is a pheromone released by plants when they are attacked by aphids. It activates plant defence systems, as well as attract predators of the aphids, such as the ladybug or the green lacewing. In the lab we have focused on increasing the production of methyl salicylate of an existing brick, by increasing the production of its precursor, chorismate.

Methyl Salicylate - Modelling

We have modeled the production system of methyl salicylate by using the transcription, translation and protein degradation rate in order to calculate the mRNA and protein flux. We also brought the kinetics of methyl salicylate synthesis into account.

Methyl Salicylate - Flux Balance Analysis

We ran the Flux Balancing Analysis using the COBRA Toolbox for MATLAB. With this analysis we checked whether the growth rate of our cells will be affected when we introduce our methyl salicylate system.

qPCR

We did a qPCR to check whether our genes of interest are properly transcribed. Also, the amount of mRNA we can measure with qPCR could serve as input data for our methyl salicylate model.