Team:Paris/Transduction overview strategy

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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_strategy#bottom | Our strategy]]
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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_strategy#bottom | Our strategy]]
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== Overview  ==
 
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* [[Team:Paris/Transduction_overview#Overview | Introduction]]
 
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* [[Team:Paris/Transduction_overview_fusion |A. Fusion ]]
 
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** [[Team:Paris/Transduction_overview_fusion#junfos |A.1 jun/fos]]
 
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** [[Team:Paris/Transduction_overview_fusion#g3p |A.2 G3P]]
 
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** [[Team:Paris/Transduction_overview_fusion#snare |A.3 Snares]]
 
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* [[Team:Paris/Transduction_overview_transduction |B. Transduction]]
 
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** [[Team:Paris/Transduction_overview_transduction#ABCT |B.1 ABC transporters]]
 
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** [[Team:Paris/Transduction_overview_transduction#TCS |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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===C. Our Strategy===
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==Membran Fusion : Our Strategy==
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<a class="menu_sub"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_active"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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====C.x Fec====
 
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Few ABC transporter such as FecABCD (iron transporter) are able to induce a response regardless of the tranlocation, due to the activity of FecA, some mutant can also have a constitutive expression of FecABCD . We could use FecA- mutant  receiver
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===Jun/Fos strategy===
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and FecA+ mutant donor to transfert working FecA to the receiver. In this case the receiver will express the FecABCD operon, and so we could place under the control of the Fec ABCD promoter the gene sequence encoding for the response.
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Our strategy is to fuse the mutated Jun (the one unable to form homodimer) to AIDA, the autotransporter (for the donnor cell). By this way, Jun will be export to the outer membrane and should be on vesicles.
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The second step is to make a fusion between Fos and AIDA (for the receiver bacteria). In this direction Fos will be trasport to the outer membrane .
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===g3p strategy===
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g3p is naturally exposed at the surface of the filamentous bacteriophage. 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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[[Image:Transduction_overview_fec.png|500px|center|thumb| fec operon induction]]
 
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Outer membrane protein A (OmpA) is a major structural protein of the outer membrane of Escherichia coli.
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'''problems :''' the message is unidirectionnal and unrepeatable. It would just be a proof of principle that a vesicle-mediated controlled communication is possible.
 
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====C.x The trick TCS====
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To conclude, our strategy is to fuse OmpA-Linker to g3p to export the g3p protein to the outer membrane in order to find them on the vesicles surface.
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We could with the help of Alfonso Jaramillo design a synthetic PBP which could detect the substrate we choosed and activate a specific HK.
 
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The previous work (Valencia 2006) design a vanilin-sensitive PBP and a network for a graduated response whereas we just need a proteic sensitive PBP and a binary type of response.
 
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strain and part needed:
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{{Template:Paris2009_guided|Transduction_overview_fusion#bottom.23top|Transduction_overview2#top}}
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strain : E.coli MRi7 (unable to translocate ribose from periplasm to cytoplasme)
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part :
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Latest revision as of 22:41, 21 October 2009

iGEM > Paris > Receiving the message > Our strategy


Membran Fusion : Our Strategy


Jun/Fos strategy

Our strategy is to fuse the mutated Jun (the one unable to form homodimer) to AIDA, the autotransporter (for the donnor cell). By this way, Jun will be export to the outer membrane and should be on vesicles. The second step is to make a fusion between Fos and AIDA (for the receiver bacteria). In this direction Fos will be trasport to the outer membrane .

g3p strategy

g3p is naturally exposed at the surface of the filamentous bacteriophage. 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.


Outer membrane protein A (OmpA) is a major structural protein of the outer membrane of Escherichia coli.


To conclude, our strategy is to fuse OmpA-Linker to g3p to export the g3p protein to the outer membrane in order to find them on the vesicles surface.


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