Team:Oxford/biosensor construction

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Construction


In order to be able to use our model and to determine whether DcmR acts as a repressor or activator in the presence of DCM we designed the following two plasmid systems:
pOXON-2-dcmR-mCherry
pOXON-2-dcmR-mCherry
Oxford iGEM 2014
pSRK-Gm-pdcmAsfGFP
pSRK-Gm-pdcmAsfGFP
Oxford iGEM 2014
Why these two plasmid backbones?
Why these two plasmid backbones?
  • The two plasmids are partitioned during cell division by different systems, thus an equal proportion of each plasmid is maintained in each new daughter cell.

  • Different antibiotic resistances will allow us to select for cells that have taken up both plasmids by application of both antibiotics.

  • The replication origins compatible with E.coli and pseudomonas strains.

  • We have used two plasmids so that we can test each part in isolation before transforming them both into the same cell.
    • How where the constructs made?
    How where the constructs made?
  • The two plasmids are partitioned during cell division by different systems, thus an equal proportion of each plasmid is maintained in each new daughter cell.

  • Different antibiotic resistances will allow us to select for cells that have taken up both plasmids by application of both antibiotics.

  • The replication origins compatible with E.coli and pseudomonas strains.

  • We have used two plasmids so that we can test each part in isolation before transforming them both into the same cell.