35S is a plant specific 3' untranslated sequence and terminator from Cauliflower Mosaic Virus. The part’s intended use is to terminate transcription in plant cells. This part was converted to a standard BioBrick by cloning into the Mo-Flipper Plasmid K1467300 made by the NRP-UEA 2014 team.
NRP-UEA 2015 team used this part in our plant constructs. Constructs K1618029-36 contained a 35s promoter, chloroplast transit peptide, one of four acyltransferases, and this new 35s terminator parts. In parts K1618029-32 fused the recombinant protein was fused to a fluorescent a reporter tag to confirm the localisation to the chloroplast (Figure 1). The parts were tested by transferring into Agrobacterium tumefaciens and infiltration into plants. The correct and strong expression indicates that the terminator is functional.
Figure 1: Constructs BBa_K1618029‐032 contain a yellow fluorescent protein, as well as a chloroplast transit peptide. These are confocal microscopy images of the constructs infiltrated into Nicotiana benthamiana, in which the red structures are the chlorophyll within the chloroplast, and the yellow is the fluorescent fusion protein expressed from constructs (a) BBa_K1618029, (b) BBa_K1618031, (c) BBa_K1618032, and (d) BBa_K1618030.
The above results not only indicate a successful localisation, but also suggest that the 35s terminator that we used is GoldenGate compatible and functioning.