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Team:Paris/Project
From 2009.igem.org
Overall project: Message in a bubble
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Plasmid construction
To implement our vesicles emission project, we had to think about a few constraint in order to make our system efficient. To put into place all the functionalities we needed, we decided to use 2 different plasmids.
First of all, before sending vesicules into the surrounding medium, we have to make sure that every molecule and protein that has to be inside the vesicules is already into place before the bacteria starts the creation of vesicules. In other words, the "emitting" bacteria starts producing the protein of interest, the export systems, the FecA proteins as well as the fusion mechanism before creating vesicules.
To create this delay between the creation of proteins and the production of vesicules, we used a double repressor made of LacI and TetR. The LacI biobrick in placed in plasmid A, downstream the pBad promoter and once it is synthetised, it acts as a repressor on the pLac promoter in plasmid B thus inhibiting the production of TetR. The ptet promoter is no longer repressed by TetR and the creation of non functional TolR can start as well as the emission of vesicules.
As the creation of vesicules via the over expression of TolR
Modeling Overview
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quoi que il se peut que certaines pages soient remplies tranquillement ... ou pas
