Team:Paris/Production design

From 2009.igem.org

Revision as of 13:15, 8 September 2009 by Cambray (Talk | contribs)

iGEM > Paris > Production > Parts Design

Contents

Parts

Registered parts we used

Backbones

The function "increased production of OMV" is borne on two plasmid backbones pSB1A3 and pSB3T5.

Biobricks

  • Expression of the proteins targeted to OMVs as well as lacI is controled by the Pbad promoter BBa_K11309.

We used the Ptet promoter BBa_ROO40 and the coding sequence for the TetR repressor protein 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 BBa_B0014 was used to isolates the different expression units from each other.

Our contributions

  • clyA fusion N-term and C-term

NB: All coding sequences intented to be expressed are associated with a standard RBS incorporated in the forward primer used to amplify the sequence by PCR. The standard RBS used was carefully designed to ensure maximal functionality (see xxx)


The RBS were designed by Guillaume Cambray and integrated directly inside the primers.