Team:Paris/Addressing overview strategy

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<span/ id="bottom">[https://2009.igem.org/ iGEM ] > [[Team:Paris#top | Paris]] > [[Team:Paris/Addressing_overview#top | Adressing]] > [[Team:Paris/Addressing_overview#bottom | Export & ClyA]] > [[Team:Paris/Addressing_overview_strategy#bottom | Our strategy ]]
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<span/ id="bottom">[https://2009.igem.org/ iGEM ] > [[Team:Paris#top | Paris]] > [[Team:Paris/Addressing_overview#top | Adressing]] > [[Team:Paris/Addressing_overview#bottom | Export systems]] > [[Team:Paris/Addressing_overview_strategy#bottom | Our strategy ]]
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== Overview ==
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* [[Team:Paris/Addressing_overview#Overview | Introduction]]
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* [[Team:Paris/Addressing_overview_exportsystem#Overview |A. Export system ]]
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==Adressing the message to the cytoplasm : Our strategy==
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* [[Team:Paris/Addressing_overview_clya#Overview |B. ClyA ]]
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* [[Team:Paris/Addressing_overview_strategy#Overview |C. Our strategy]]
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* [[Team:Paris/Addressing_overview_Construction#Overview |D. Construction]]
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===C. Our strategy===
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<a class="menu_sub" href="https://2009.igem.org/Team:Paris/Addressing_overview_exportsystem#bottom"> Export system</a>|
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<a class="menu_sub_active"href="https://2009.igem.org/Team:Paris/Addressing_overview_strategy#bottom"> Our Strategy</a>|
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ClyA:
 
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Our strategy is in the first time link the RFP fusion to ClyA-Nterm, we can also link it in ClyA-Cterm but it seems to be lee visible when using microscopy to observ the visicle. We also have to put the promoter pbad forward the ClyA RFP fusion.  
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We thought of overexpressing the important proteins in the Tat pathway (that is to say TatABCE) in order to avoid the early saturation phenomenon that is likely to occur when we will overexpress proteins that will be targeted to the outer membrane or to the periplasm.
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This construction is maked in order to show that we can put some protein in vesicle and transport him to its target cell, so when it'll work we will fusion it with a signal peptide for the signal transduction.
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However, in our strategy, the only protein that needs to use the TAT pathway to translocate from the cytoplasm to the periplasm is clyA. Finally, we decided that the overexpression won't be necessary neither for the TAT pathway, nor for the SEC pathway, both of them constitutively expressed in E Coli K12.

Latest revision as of 06:47, 21 October 2009

iGEM > Paris > Adressing > Export systems > Our strategy


Adressing the message to the cytoplasm : Our strategy


We thought of overexpressing the important proteins in the Tat pathway (that is to say TatABCE) in order to avoid the early saturation phenomenon that is likely to occur when we will overexpress proteins that will be targeted to the outer membrane or to the periplasm.

However, in our strategy, the only protein that needs to use the TAT pathway to translocate from the cytoplasm to the periplasm is clyA. Finally, we decided that the overexpression won't be necessary neither for the TAT pathway, nor for the SEC pathway, both of them constitutively expressed in E Coli K12.

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