Team:Paris/Production overview strategy

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iGEM > Paris > Production > Overview > Our Strategy

Overview

B. Our strategy

Note de Stoff (je sais je suis chiant mais c'est qu'un début) : Il faut développer le choix de Tol/pal :

  • faire un liens l'inventaire des publications qui disent que ca existe et que c'est bien foutu et 'connu' (en gros la partie d'avant présentation de Tol/pal) + liste des équipes iGEM qui ont/vont travailler dessus de près ou de loin.
  • faire l'inventaire des (de là) publication(s) qui a/ont prouvé que ca marche (ou pas :d). (proposition d'aide (liens vers nos amis... ou pas).
  • en conclure que connaissant les autres system celui là allait bien dans notre choix car : connu, facilement faisable dans le temps qui nous était imparti, obiwan kenobi toussa toussa

The first strategy is the knock out of tol/pal gene witch destabilize the membrane.It's an active system, whereas the use of conditional mutant is a passive system. A previous study[2] focused on the development of a gene expression system able to induce production of large amounts of OMVs.


To achieve our goal, we decided to take advantage of the second domain of TolR (TolRII). We don’t use the third domain of TolA because, it doesn’t work so good and TolAIII have lot of PstI domain in his sequence. In order to achieve our project, we will over express specifically designed biobricks containing TolRII fused with OmpA signal which allows it to migrate in the periplasm[3].


In the same framework, we could also over express various Tol ligand (like colicin) to destabilize the membrane.


References

  1. ^The Tol-Pal proteins of the Escherichia coli cell envelope an energized system required for outer membrane integrity. Lloubès & Journet 2001 - [http://www.ncbi.nlm.nih.gov/pubmed/11501670 11501670]
  2. ^Improved methods for producing outer membrane vesicles in gram-negative bacteria. Henry & Lloubès 2004 - [http://www.ncbi.nlm.nih.gov/pubmed/15249060 15249060]

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