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Team:UNITN-Trento/Project/Datapage
From 2013.igem.org
We envision this genetic circuit in which production of ethylene and methyl-salycilate (MeSA) are regulated by light. In a dark state, the sensor protein YF-1 auto-phosphorilates itself, dimerize, and transfer the phosphate to the response regulator element FixJ. FixJ is so activated and is able to bind pFixK2 promoter, activating transcription of cI (inhibitor protein) that ultimately blocks EFE production. At the same time, it is transcribed Bmst1, which regulates MeSA synthesis thus slowing down fruit ripening. In a blue-light state the sensor protein YF-1 does not phosphorilate itself, so pFixK2 promoters is in the off state: ethylene is produced thus promoting ripening while MeSA production is blocked.
These are our best charachterized parts on the system
- Best natural part: BBa_K1065000, 2-Oxoglutarate Oxygenase/Decarboxylase Ethylene Forming Enzyme (EFE). That part was well characterized under the control of different inducible promoters in both E. coli (NEB10beta cells) and in B. subtilis (str.168).
- Best device: BBa_K1065310, blue light regulated AmylCP producing device. This device includes an iverter cassette (composed by cI protein and pLambda promoter) that ultimately allows production of the chromoprotein AmilCP only when the culture is exposed to light.
- Best improved part: BBa_K1065302, blue light regulated AmilGFP producing device (a yellow fluorescent protein). The part BBa_K952003 was improved inserting an RBS sequence between pFixK2 promoter and AmilGFP coding sequence by mutagenesis. We demonstrated than that our part produced successfully AmilGFP when the culture was mantained in the dark.
