Team:Paris/Addressing overview strategy

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== Overview ==
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* [[Team:Paris/Addressing_overview_exportsystem#Overview |A. Export system ]]
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* [[Team:Paris/Addressing_overview_strategy#Overview |B. Our strategy]]
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==Adressing the message to the cytoplasm : Our strategy==
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* [[Team:Paris/Addressing_overview_Construction#Overview |C. Construction]]
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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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<a class="menu_sub"href="https://2009.igem.org/Team:Paris/Addressing_testing#bottom"> Construction</a>
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==B. Our strategy==
 
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Our strategy is to use clyA to export a protein to the outer-membrane of the cell. The protein fused to clyA will be incorporated into the vesicle during the vesiculation process. ClyA contain the signal peptide needed to be exported form the cytoplasm to the periplasm. The overall idea is to fused the protein of interest to clyA. So, to make sure that there isn't an early saturation phenomenon which will disturb the equilibrium between the vesiculation process and the protein translocation to the periplasm. That's the reason why we thought of overexpressing the important proteins in the Tat pathway (that is to say TatABCE) in order to avoid this early saturation phenomenon. 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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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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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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