Difference between revisions of "Part:BBa K382021"

Revision as of 15:12, 5 November 2010

Arabidopsis optimized Miraculin

Miraculin is a 'flavor inverting' protein, found naturally in the fruit of the plant fruit of Synsepalum dulcificum. Not sweet by itself, miraculin binds to taste receptors on the tongue, possibly altering the structure of the receptors and causing traditionally 'sour' flavors to be received as 'sweet'.

BBa_K382021 is an Arabidopsis codon-optimized version of miraculin. We characterized expression of YFP-tagged versions in E. coli. The C-terminal 2xYFP tagged version has been submitted to the registry (BBa_K382040).

The graphs below show IPTG-induction of miraculin over time in E. coli liquid cell culture (red line) compared to uninduced cultures (blue line). 2xYFP tags were attached to either N- (panel A) or C- terminus (panel B) of miraculin to ensure that YFP position had no influence over expression. In all cases relative miraculin expression (indicated by YFP fluorescence) had appreciably increased after 120 minutes as compared to the non-induced E. Coli.

Confirmed function: We have confirmed that the BioBricked Arabidopsis codon-optimized miraculin protein can be tagged with YFP and expressed from am IPTG inducible BioBrick promoter in E. coli. Due to safety restrictions, the flavor inverting function of miraculin expressed from E. coli has not been tested. Continued work will allow us to produce Miraculin in form that is safe for taste testing.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

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 Miraculin is a 'flavor inverting' protein, found naturally in the fruit of the plant fruit of Synsepalum dulcificum. Not sweet by itself, miraculin binds to taste receptors on the tongue, possibly altering the structure of the receptors and causing traditionally 'sour' flavors to be received as 'sweet'.
-We characterized[http://2010.igem.org/Team:Harvard/flavor/results] expression of the YFP-tagged version of this part in ''E. coli''. [[Part:BBa_K382040|BBa_K382040]]
+BBa_K382021 is an Arabidopsis codon-optimized version of miraculin. We characterized expression of YFP-tagged versions in ''E. coli''. The C-terminal 2xYFP tagged version has been submitted to the registry ([[Part:BBa_K382040|BBa_K382040]]).
-xYFP tags were attached to either N- (panel A) or C- terminus (panel B) of miraculin 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.
+The graphs below show IPTG-induction of miraculin over time in E. coli liquid cell culture (red line) compared to uninduced cultures (blue line). 2xYFP tags were attached to either N- (panel A) or C- terminus (panel B) of miraculin to ensure that YFP position had no influence over expression. In all cases relative miraculin expression (indicated by YFP fluorescence) had appreciably increased after 120 minutes as compared to the non-induced E. Coli.
-YFP tag performance:
 <br>
 [[Image:Mir_BrazzYFP_Fig_1-crop.jpeg|600px]]
-Conclusion: We have confirmed that the BioBricked miraculin protein can be tagged with YFP and expressed from am IPTG inducible BioBrick promoter in E. coli. The flavor inverting function of miraculin expressed from E. coli has not been teasted, due to safety restrictions. Continued work will allow us to produce Miraculin in form that is safe for consumption.
+Confirmed function: We have confirmed that the BioBricked Arabidopsis codon-optimized miraculin protein can be tagged with YFP and expressed from am IPTG inducible BioBrick promoter in E. coli. Due to safety restrictions, the flavor inverting function of miraculin expressed from E. coli has not been tested. Continued work will allow us to produce Miraculin in form that is safe for taste testing.