Results
![]()
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. 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
|
Confirmation with Western Blot
A western blot assay was performed to check for E. Coli expression of
|
Figure 2. Western Blot of Miraculin and Brazzein Expression in E. Coli
Proteins were tagged using a StrepII standard Antibody tag, attached to both N- and C- termini. Miraculin (A) does not appear to have been expressed in high enough quantities to be visualized. The expected protein weight is 25 kDa. Brazzein (B) shows strong expression of a protein in the 10-15 kDa range. Brazzein has an expected weight of 6.5 kDa, a discrepancy that we have attributed to inconsistencies in the gel.
|
"
