Team:ASIJ TOKYO/Results
Results
Overview
We attached the COX-2 (BBa_K2226003) and c-Myc (BBa_K2226002) promoters to the half of the reporter systems that yielded the best results. We were inspired by the 2015 Peking iGEM team to use nonspecific binding sites FRB and FKBP. (Peking iGEM Team 2015, 2015) (Rossi, Charlton, & Blau, 1994) (Rem, Galarneau, & Michnick, 2001). These reporter systems included the rapamycin-binding domain (FRB) fused to nLuc and the FK-506 binding protein 12 (FKBP) fused to cLuc. The fusion of the promoter to the reporter system made the nonspecific binding sites on the domains to become specific to the COX-2 (BBa_K2226003) and c-Myc (BBa_K2226004) proteins (Tomano & Nobuyuki, 2016). The newly specialized promoter-reporter system would then only come together in the presence of these two proteins. The creation of the FRB-rapamycin-FKBP complex induced the fusion of nLuc and cLuc, in turn generating light in the presence of rapamycin (Peking iGEM Team 2015, 2015).
Promoter Strength Confirmation
Construction of the Promoter-Reporter System
Proof of Concept
Confirmation
