Team:TU-Eindhoven/Parts

iGEM TU Eindhoven

Parts

We, the iGEM team from Eindhoven University of Technology, created eight BioBrick parts that show the functionality and orthogonality of a new scaffold platform. The four scaffold parts are T14-3-3 heterodimers. The other four parts consist of a target protein attached to the free N-terminus of the CT52. Under influence of the small molecule fusicoccin, two different protein-CT52 complexes (two target proteins) can dimerize on the scaffold. The binding between T14-3-3 and CT52 is reversible when fusicoccin is removed.

NameTypeDescriptionDesignerLength
   BBa_K2065000CodingSmallBiT-CT52Tom van Sonsbeek294
  BBa_K2065001CodingmNeonGreen-CT52Tom van Sonsbeek969
  BBa_K2065002CodingT14-3-3 S71L – T14-3-3 scaffoldTom van Sonsbeek1548
  BBa_K2065003CodingCaspase 9-CT52Tom van Sonsbeek 
  BBa_K2065004CodingT14-3-3 (W237R) - T14-3-3Tom van Sonsbeek1548
  BBa_K2065005CodingT14-3-3 - T14-3-3 (E19R)Tom van Sonsbeek1548
  BBa_K2065006CodingT14-3-3 (S71L/I72V) - T14-3-3Tom van Sonsbeek1548
  BBa_K2065007CodingLargeBiT - CT52Tom van Sonsbeek735

We have improved two biobricks from iGEM team TU Eindhoven 2015, namely the two NanoBiT fragments called SmallBiT BBa_K1761006 and LargeBiT BBa_K1761005. We have expanded on their characterisation and application possibilities by linking it to our CT52 protein and documented our improvements on their main pages. SmallBit and LargeBit, attached to CT52, offers the ability to test a scaffold protein or to verify the working of newly found mutations in a scaffold protein.

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