Team:Paris/Transduction overview

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https://2009.igem.org/Team:Paris/Transduction_overview#top
https://2009.igem.org/Team:Paris/Transduction_testing_fusion

Overview

• Introduction
• A. Fusion
  • A.1 jun/fos
  • A.2 G3P
  • A.3 Snares
• B. Our strategy
• C. Construction

Introduction

This part of the project was focus on a precise point

• A. The fusion between the OMVs and the targeted bacteria.

We have planned to explore three different method :

With the Jun/Fos dimere: Jun and Fos are able to form an heterodimer which has a high stability and Jun can dimerize with another Jun (thanks to their leucine zipper motif). After mutations into the leucine zipper motif of Jun (that allow the Jun/Fos dimerization but avoid the Jun/Jun homodimer formation), we wanted to fuse it to AIDA (an ABC transporter) to send them to the extern membrane of bacteria.

With G3P : The viral protein known as G3P is naturally exposed at the surface of the filamentous bacteriophage which enable it to get in the bacteria. The M13 phage has a high affinity for E.coli, and if we could place its G3p on the surface of the vesicles it could activate the fusion with the Outer membrane of the targeted bacteria.

In order to target the G3P at the surface of of the vesicles, we fuse it to the OmpA- Linker protein (created by the Warsaw team)

With the SNARE system:

Intracellular membrane fusion in eukaryotes requires SNARE (soluble N-ethylmaleimide-sensitive-factor attachment protein receptor) proteins that form complexes bridging the two membranes. To allow this fusion, the perfect conformation of all protein that composed the SNARE complex is an obligation. Untill now, no one cloned this complex into bacteria. We thank that it will be a very "dangerous way" to go for our project, so we decided to focus our effort on the Jun/Fos strategy and the G3P one.