FRB/FKBP control targetSCREENER assay
The rapamycin-inducible interaction between a membrane-bound FRB domain (FKBP-binding domain of mTOR) (TM-FRB) and a cytosolic FKBP is a very sensitive targetSCREENER control for the splitSENSOR technology (Figure 1). In this assay, TM-FRB is fused to the NTEV moiety, alongside a TEV protease cleavage site (tevS) and the artificial transcriptional co-activator Gal4-VP16 (GV); FKBP is fused to CTEV. Both the TM-FRB-NTEV-tevS-GV fusion (driven by a CMV promoter) and the FKBP-CTEV fusion (driven by an IRES) are assembled on one plasmid allowing a robust and simple assay setup as only the final UAS-driven reporter (i.e. firefly luciferase) needs to be added.
Figure 1: The targetSCREENER control assay. The rapamycin-inducible interaction between membrane-bound FRB (TM-FRB) and cytosolic FKBP was monitored in a dose response assay. The EC50 value is at 0.9nM.
To order the FRB/FKBP control targetSCREENER assay, simply select from the two options available in the section product listings. The first option contains the pIndControl plasmid (SYS-V529) only, the second option contains both pIndControl and the firefly reporter gene plasmid pG10_Fluc (targetSCREENER control set, SYS-S01).
Please note that an annotated sequence for each plasmid will be made available once it has been ordered.