Difference between revisions of "Part:BBa K382020"

Revision as of 15:38, 5 November 2010

Arabidopsis optimized Brazzein

Brazzein is a sweet-tasting protein that is found in the Western African fruit, the Oubli (Pentadiplandra brazzeana). The protein consists of a 54 amino acid sequence. It is sweeter than sugar and is used as an alternative low calorie sweetener.

We characterized[http://2010.igem.org/Team:Harvard/flavor/results] expression of the YFP-tagged version of this part in E. coli. BBa_K382041

BBa_K382020 is an Arabidopsis codon-optimized version of brazzein. We characterized expression of YFP-tagged versions in E. coli. The C-terminal 2xYFP tagged version has been submitted to the registry (BBa_K382041). 2xYFP-tagged miraculin was expressed from a IPTG-inducible Novagen Duet vector that was adapted to the BioBrick standard (described in [http://www.jbioleng.org/content/4/1/3 Agapakis et al., JBE, 2010]).

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 C) or C- terminus (panel D) 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.

Note: Panels (A) and (B) show similar characterization of another flavor protein, Arabidopsis codon-optimized miraculin.

YFP tag performance:

We also characterized this part with N- and C-terminal strepII tags, using the same IPTG-inducible Novagen Duet vector system. Strep tag performance:

Sequence and Features