Team:Paris/Addressing overview strategy

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iGEM > Paris > Adressing > Export systems > Our strategy

Overview

B. Our strategy

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


In fact 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 we thought that it could be necessary to overexpress TatABCE .

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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