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Team:Paris/Production overview strategy
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==Overview== | ==Overview== | ||
| - | + | * [[Team:Paris/Production_overview#Overview | Introduction]] | |
| + | * [[Team:Paris/Production_overview_Tol/Pal#Overview |A. Tol/pal ]] | ||
| + | * [[Team:Paris/Production_overview_strategy#Overview |B. Our strategy]] | ||
| + | * [[Team:Paris/Production_overview_Construction#Overview |C. Construction]] | ||
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Revision as of 18:27, 31 August 2009
iGEM > Paris > Production > Overview > Our Strategy
B. Our strategy
The first strategy is the knock out of tol/pal gene witch destabilize the membrane .It is 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 overexpress 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 overexpress various Tol ligand (colicin) to destabilize the membrane.
References
[1] Transmembrane a-helix interactions are required for the functional assembly of the escherichia coli Tol complex. Lazzaroni & Geli 1995
[2] The Tol-Pal proteins of the Escherichia coli cell envelope an energized system required for outer membrane integrity. Lloubès & Journet 2001
[3] Improved methods for producing outer membrane vesicles in gram-negative bacteria. Henry & Lloubès 2004
