Team:NRP-UEA-Norwich/Collaborations/Valencia

UPV Valencia collaboration

As the Valencia (UPV) team are also working on plants and with Type IIS-assembly methods (RFC 106) we collaborated by sending each other plant-expression constructs.

We sent Valencia four of our plant parts (BBa_K1618029, BBa_ K1618030, BBa_ K1618031, and BBa_ K1618032) all of which contained a YFP fluorescent reporter tag. We also sent them the ‘MoClo Flipper Constructs’ (BBa_K1467100, 200, 300 and 400) that were made by the 2014 NRP-UEA team, to allows them to easily convert their GoldenGate parts to standard Biobricks for submission to the registry.

They sent us two parts (a) 35s; T35S – p.ReporterEscission; GFP; T35s and (b) ReporterEscission;GFP;T35s so that we could determine the subcellular localisation of GFP in a different plant chassis to the one that they were using.

We transformed their parts into Agrobacterium tumefaciens according to the Agro-transformation protocol (described on the Protocols page). We then infiltrated Nicotiniana tabacum according to the Agro-infiltration protocol (described on the Protocols page) and, after two days, imaged the leaves on a confocal microscope (Figure 1). Fluorescence was observed in the cytoplasm as well as in the nucleus from both constructs. Our results were consistent with the results that they obtained in Nicotiana benthamiana.

Figure 1: The confocal microscope images showed GFP fluorescence in the nucleus and cytoplasm of Nicotiana tabacum cells for both (a) 35s; T35S – p.ReporterEscission; GFP; T35s and (b) ReporterEscission;GFP;T35s. This is consistent with what the Valencia team observed in Nicotiana benthamiana.