Team:Wageningen UR/Notebook/OptogeneticKillSwitch

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Wageningen UR iGEM 2016

 

These experiments were performed primarily by Marijn, with significant contributions by Mark during the final phase. Unless otherwise indicated, protocols were done according to the Protocols page.

May 16 - June 6: Obtaining Parts

Both MazE and MazF have been registered as BioBricks (parts BBa_K1096001 and BBa_K302033, respectively). However, as of the creation of the kill switch, neither are available from the iGEM registry. For this reason, we chose to isolate both genes directly from E. coli genomic DNA via PCR. In addition, the following BioBricks were prepared from iGEM registry kits:

  • BBa_K592006: FixK2 (Promoter triggered by YF1-FixJ)
  • BBa_K592016: YF1-FixJ (Light sensor)
  • BBa_B0015: Double terminator
  • BBa_J23108: Constituitive promoter of the Anderson family (Mid-level expression strength, arbitrarily chosen.)

    • 4 July - 1 August: pDawn Construction

    8 August - 3 October: pDusk Construction