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Confirmation with YFP-2x Tags

In order to confirm that the Miraculin and Brazzein are able to be expressed in E. Coli we attached a YFP-2x tag sequence to the termini of both proteins. The proteins were placed under an IPTG-expressible promoter and used spectrophotometry to determine the level of YFP fluorescence against a baseline, untagged protein. Figure 1 shows relative-fluorescence at times post induction. In all circumstances the levels of YFP-fluorescence increased

Figure 1   click to enlarge

Figure 1. Induced expression of YFP-tagged Miraculin and Brazzein in E. Coli Figure 1 (A) through (D) are normalized plots of Miraculin and Brazzein YFP-fused constructs expressed in E. Coli. YFP-2x tags were attached to both N- and C- termini to ensure that folding was not hindered. In all cases relative YFP fluorescence had appreciably increased after 120 minutes as compared to the non-induced E. Coli