Team:Paris/Transduction overview

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<span/ id="bottom">[https://2009.igem.org/ iGEM ] > [[Team:Paris#top | Paris]] > [[Team:Paris/Transduction_overview#top | Reception]] > [[Team:Paris/Transduction_overview#bottom | Overview]]
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<span/ id="bottom">[https://2009.igem.org/ iGEM ] > [[Team:Paris#top | Paris]] > [[Team:Paris/Transduction_overview#top | Receiving the message]] > [[Team:Paris/Transduction_overview#bottom | Membrane fusion]]
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== Overview  ==
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* [[Team:Paris/Transduction_overview#Overview#bottom | Introduction]]
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* [[Team:Paris/Transduction_overview_fusion#bottom |A. Fusion ]]
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** [[Team:Paris/Transduction_overview_fusion#A.1 Jun/Fos|A.1 jun/fos]]
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==Membrane fusion: Main==
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** [[Team:Paris/Transduction_overview_fusion#A.2 G3P|A.2 G3P]]
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** [[Team:Paris/Transduction_overview_fusion#A.3 Snares |A.3 Snares]]
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* [[Team:Paris/Transduction_overview_transduction#bottom |B. Transduction]]
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** [[Team:Paris/Transduction_overview_transduction#B.1 ABC transporters|B.1 ABC transporters]]
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** [[Team:Paris/Transduction_overview_transduction#B.2 Two-component system|B.2  Two Component System]]
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* [[Team:Paris/Transduction_overview_strategy#bottom |C. Our strategy]]
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* [[Team:Paris/Transduction_overview_construction#bottom |D. Construction]]
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<a class="menu_sub_active"href="https://2009.igem.org/Team:Paris/Transduction_overview#bottom"> Main </a>|
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<a class="menu_sub"href="https://2009.igem.org/Team:Paris/Transduction_overview_fusion#bottom"> Fusion</a>|
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<a class="menu_sub"href="https://2009.igem.org/Team:Paris/Transduction_overview_strategy#bottom"> Our strategy</a>|
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<a class="menu_sub"href="https://2009.igem.org/Team:Paris/Transduction_overview_construction#bottom"> Construction</a>
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===Introduction===
 
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- to enable gene transcription after fusionning OMVs with the outer membrane of the receiving bacterium.
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This part of the project was focus on a precise point
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*[[Team:Paris/Transduction_overview_fusion#bottom |The fusion between the OMVs and the targeted bacteria.]]
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We also hope that we could achieve this aim without sacrifying important proprieties of our message : specific, repeatable , multidirectional.
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We have planned to explore three different method :
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It seems that we have two possible ways : the ABC transporters or the two component systems .
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ABC transporters and two component systems are natural transport system (export or import) of information, nutriments or toxines.
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'''''With the Jun/Fos dimere:'''''
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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).
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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.
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'''''With G3P :'''''
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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.
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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)
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There is another possibility but the mecanism is mostly unknown : the DNA-containing OMVs which could be a useful mean of information transport.
 
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====References====
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'''''With the SNARE system:'''''
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<ol class="references">
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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 thought 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.
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<li> [[Team:Paris/Transduction_overview#1 | ^]] - [http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=179564 179564]</li>
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<li> [[Team:Paris/Transduction_overview#2 | ^]]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]</li>
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<li> [[Team:Paris/Transduction_overview#3 | ^]]Improved methods for producing outer membrane vesicles in gram-negative bacteria. Henry & Lloubès 2004 - [http://www.ncbi.nlm.nih.gov/pubmed/15249060 15249060]</li>
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#1995- Cosima Harle & Volkmar Braun - Signal transfer through three compartments transcription initiation of the Escherichia coiferric  citrate transport system from the cell surface
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{{Template:Paris2009_guided|Production_overview#bottom|Transduction_overview_fusion#bottom}}
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#2000 - Stock & Goudreau – Two-component signal  transduction
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#2001 - Mishima & Murata - Super-channel in bacteria  function and structure of the macromolecule import system mediated by a pit-dependent ABC transporter
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#2009 - Akyriakidis & Tiligada – signal transduction TCS  the AtoSC paradigm
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#2009 - Tomii & Kanehisa – comparative analysis of ABC transporter
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Latest revision as of 15:06, 21 October 2009

iGEM > Paris > Receiving the message > Membrane fusion



Membrane fusion: Main


This part of the project was focus on a precise point

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 thought 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.


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