Team:Harvard/flavor/results
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In order to confirm that the Miraculin and Brazzein are able to be expressed in <i>E. Coli</i> we attached a <i>YFP-2x</i> 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 | In order to confirm that the Miraculin and Brazzein are able to be expressed in <i>E. Coli</i> we attached a <i>YFP-2x</i> 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 | ||
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+ | <div>reporter series <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/> | ||
+ | <a href="https://static.igem.org/mediawiki/2010/7/77/YFP_Fig_1-crop.jpg" id="single_image"> | ||
+ | <img src="https://static.igem.org/mediawiki/2010/7/77/YFP_Fig_1-crop.jpg" width="300px" border=0> | ||
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+ | <td style="vertical-align:top"><p style="padding:10px"><br/>The reporter series vectors contain a reporter on the trailing end of the multiple cloning site such that expression of the reporter follows that of the inserted construct. We modified the vectors pORE R1 and pORE R3. pORE R1 contains the gusA reporter, and pORE R3 the smgfp reporter. Both vectors confer plant resistance to kanamycin. </p></td> | ||
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Revision as of 02:22, 25 October 2010
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Results
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Miraculin & Brazzein
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
![](https://static.igem.org/mediawiki/2010/7/77/YFP_Fig_1-crop.jpg)
The reporter series vectors contain a reporter on the trailing end of the multiple cloning site such that expression of the reporter follows that of the inserted construct. We modified the vectors pORE R1 and pORE R3. pORE R1 contains the gusA reporter, and pORE R3 the smgfp reporter. Both vectors confer plant resistance to kanamycin.