Coding

Part:BBa_K1113701:Experience

Designed by: Belen Cespedes   Group: iGEM13_UC_Chile   (2013-09-18)

We test the targeting sequence of the RuBisCO-GFP (the large and short subunit) in a co-transformation with the Carboxysome biobick (Bba_K1113100)+ RFP, all this in the TOP10 E. coli strain, then we observe this bacterias with the confocal microscope and look for colocalization of both GFP (targeting sequence) and RFP (carboxysomes).

Team UC Microscopioconfocal.jpg
Figure 1. Confocal Microscopy. Brightfield, GFP filter, RFP filter and merge are shown.

As observed in Figure 1, two of our constructs display co-localization of green and red fluorescence. Finally we quantify the amount of colocalization of our targeting sequence and obtained (Table 1) that the construct that had the grearest co-localization was C.LG, which is the large subunit of RuBisCO with GFP attached to the carboxil terminal end of the enzime.
Nuevaimagen.jpg
Table 1. Quantify of the colocalization.

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