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Team:Paris/Addressing overview2 strategy
From 2009.igem.org
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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. | 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 in a first time in order to see if our ClyA are in OMVs, we fused it we a RFP, for that we add a poly glycine linker to ClyA biobrick design for improving ClyA-RFP fusion. Moreover if we put RFP before ClyA, it seem that there is more fluorescence than ClyA before RFP under [article 1]. You can see the contruction here [[ | + | So in a first time in order to see if our ClyA are in OMVs, we fused it we a RFP, for that we add a poly glycine linker to ClyA biobrick design for improving ClyA-RFP fusion. Moreover if we put RFP before ClyA, it seem that there is more fluorescence than ClyA before RFP under [article 1]. You can see the contruction here [[https://2009.igem.org/Team:Paris/Addressing_overview_Construction#Overview]] |
Then if this test we could replace the RFP by the protein of interest for signal transduction. | Then if this test we could replace the RFP by the protein of interest for signal transduction. | ||
Revision as of 20:47, 19 October 2009
iGEM > Paris > ClyA > Our strategy
B. Our strategy
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 in a first time in order to see if our ClyA are in OMVs, we fused it we a RFP, for that we add a poly glycine linker to ClyA biobrick design for improving ClyA-RFP fusion. Moreover if we put RFP before ClyA, it seem that there is more fluorescence than ClyA before RFP under [article 1]. You can see the contruction here [[1]]
Then if this test we could replace the RFP by the protein of interest for signal transduction.
