Team:UNITN-Trento/Project/Datapage

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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 promoter is in the off state: ethylene is produced thus promoting ripening while MeSA production is blocked.
These are our best characterized parts on the system
  • Best natural part:
  • BBa_K1065002, 2-Oxoglutarate Oxygenase/Decarboxylase Ethylene Forming Enzyme (EFE). This part was well characterized under the control of different inducible promoters in both E. coli (NEB10beta cells) and in B. subtilis (str.168).
  • Best engineered device:
    BBa_K1065310, blue light regulated AmylCP producing device. This device includes an inverter 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_K1065305, blue light regulated AmilGFP producing device (a yellow fluorescent protein). The part BBa_K952003, extracted from the registry, was not working because it was missing the RBS between pFixK2 promoter and AmilGFP coding sequence. We improved this part by mutagenesis and demonstrated that it is able to produce AmilGFP in the dark. .
We also characterized the following parts:
  • BBa_K1065102: methyl-salycilate producing device.
  • BBa_K1065106: an improved version of our methyl-salycilate producing device. This device include the SAM synthetase gene to boost wintergreen production.
  • BBa_K1065311: EFE under the control of the blue light circuit with inverter. This device produces ethylene upon photoinduction.
Check how we exploited our parts to ripen fruit!
This is the end of the tour!
Thank you for the company!
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