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Team:Paris/Production design
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==== Biobricks ==== | ==== Biobricks ==== | ||
| - | Expression of the | + | Expression of the proteins targeted to OMVs as well as ''lacI'' is controled by the Pbad promoter [http://partsregistry.org/Part:BBa_K113009 BBa_K11309]. |
| - | We used the Ptet | + | We used the Ptet promoter [http://partsregistry.org/Part:BBa_R0040 BBa_ROO40] and the coding sequence for the TetR repressor protein [http://partsregistry.org/Part:BBa_C0040 BBa_COO40] to tightly repress the expression of protein domains that are expected to destabilize the outer membrane, thereby leading to increased production of OMVs. We hope that TetR mediated repression is strong enough to avoid leakage and weak expression of these deleterious polypeptides. |
| - | The double terminator [http://partsregistry.org/Part:BBa_B0014 BBa_B0014] was | + | The Plac promoter [http://partsregistry.org/Part:BBa_I14032 BBa_I14032] associated to ''lacI'' [BBa_C0012 http://partsregistry.org/Part:BBa_C0012] to control the expression of ''tetR''. |
| - | + | The double terminator [http://partsregistry.org/Part:BBa_B0014 BBa_B0014] was used to isolates the different expression units from each other. | |
| - | + | ||
The RBS were designed by Guillaume Cambray and integrated directly inside the primers. | The RBS were designed by Guillaume Cambray and integrated directly inside the primers. | ||
Revision as of 12:56, 8 September 2009
iGEM > Paris > Production > Parts Design
Contents |
Parts
Registered parts we used
Backbones
The function "increased production of OMV" is borne on two plasmid backbones [http://partsregistry.org/Part:pSB1A3 pSB1A3] and [http://partsregistry.org/Part:pSB3T5 pSB3T5].
Biobricks
Expression of the proteins targeted to OMVs as well as lacI is controled by the Pbad promoter [http://partsregistry.org/Part:BBa_K113009 BBa_K11309]. We used the Ptet promoter [http://partsregistry.org/Part:BBa_R0040 BBa_ROO40] and the coding sequence for the TetR repressor protein [http://partsregistry.org/Part:BBa_C0040 BBa_COO40] to tightly repress the expression of protein domains that are expected to destabilize the outer membrane, thereby leading to increased production of OMVs. We hope that TetR mediated repression is strong enough to avoid leakage and weak expression of these deleterious polypeptides. The Plac promoter [http://partsregistry.org/Part:BBa_I14032 BBa_I14032] associated to lacI [BBa_C0012 http://partsregistry.org/Part:BBa_C0012] to control the expression of tetR. The double terminator [http://partsregistry.org/Part:BBa_B0014 BBa_B0014] was used to isolates the different expression units from each other.
The RBS were designed by Guillaume Cambray and integrated directly inside the primers.
