Team:Paris/Production overview strategy

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Overview

• Introduction
• A. Tol/pal
• B. Our strategy
• C. Construction

B. Our strategy

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]