Team:Harvard/flavor/results

From 2010.igem.org

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<h1>Results</h1>
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<h2><i>YFP</i>-tagged Miraculin and Brazzein are successfully expressed</h2>
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<h2>Confirmation with <i>YFP-2x</i> Tags</h2>
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<h2>Confirmation with Western Blot</h2>
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A western blot assay was performed to check for <i>E. Coli</i> expression of
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<div>Figure 1 &nbsp; <a href="https://static.igem.org/mediawiki/2010/7/77/YFP_Fig_1-crop.jpg" id="single_image" style="font-size:12px">click to enlarge</a></div><hr/>
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<u>Figure 1. Induced expression of YFP-tagged Miraculin and Brazzein in E. Coli</u></br>
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Figure 1 (A) through (D) are normalized plots of Miraculin and Brazzein YFP-fused constructs expressed in E. Coli. <i>YFP-2x</i> 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 <i>E. Coli</i>
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Revision as of 05:06, 25 October 2010



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   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

Confirmation with Western Blot

A western blot assay was performed to check for E. Coli expression of

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

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