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.

Name	Type	Description	Designer	Length
BBa_K2065000	Coding	SmallBiT-CT52	Tom van Sonsbeek	294
BBa_K2065001	Coding	mNeonGreen-CT52	Tom van Sonsbeek	969
BBa_K2065002	Coding	T14-3-3 S71L – T14-3-3 scaffold	Tom van Sonsbeek	1548
BBa_K2065003	Coding	Caspase 9-CT52	Tom van Sonsbeek
BBa_K2065004	Coding	T14-3-3 (W237R) - T14-3-3	Tom van Sonsbeek	1548
BBa_K2065005	Coding	T14-3-3 - T14-3-3 (E19R)	Tom van Sonsbeek	1548
BBa_K2065006	Coding	T14-3-3 (S71L/I72V) - T14-3-3	Tom van Sonsbeek	1548
BBa_K2065007	Coding	LargeBiT - CT52	Tom van Sonsbeek	735

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.