Team:Nagahama/Medal Parts

• Home
• Project
  • Introduction
  • Result and Discussion
  • Future
  • Protocol
• BioBricks
  • Our all BioBriks
  • BioBriks for medal
  • PartCollection
• Attributions
  • Team
  • Attributions
  • Sponsors
• Collabolations
  • Collaborations
  • Supported by Tokyo_Tech
  • Support Tokyo_Tech
  • UGA-Georgia
  • Uniandes_Colombia
  • Paris-Saclay
  • UT-Tokyo
  • Tokyo-NoKoGen
  • UI-Indonesia
  • NEFU_China
• Human Practice
  • Human Practice
  • Risk assessment
  • Innovative HP
  • Safety
  • Sustainability of iGEM Japan
• Prototype
  • Prototype

Medal parts

銅

銀

λPL+r.b.s.+ispA＋MEP+×× (r.b.s.+dxs+r.b.s.+m-idi+r.b.s.+ispDF)

FOH is probably generated through FPP hydrolysis by endogenous phosphatases, which are induced by an increased intracellular FPP level (Asadollahi et al., 2008; Muramatsu et al., 2008a). Analogously, we hypothesized that E. coli could produce FOH under cellular conditions of an increased intracellular FPP level through metabolic engineering. A MEP pathway has been shown to synthesize IPP and DMAPP efficiently in E. coli (Martin et al., 2003). Because of its high hydrophobicity and low volatility, decane was chosen to extract and solubilize FOH from culture broth. The decane overlay in the two-phase culture did not affect growth, and FOH could be solubilized in the decane phase with negligible volatile loss. We adopt 1 mL of decane overlaid to 5 mL of culture broth. Two-phase culture of E. coli JM109 (Bba_K165025) was carried out in 2YT medium containing 1% glycerol at 29°C for 48 h. The decane phase of the two-phase culture was collected to analyze the FOH content by GC-MS. In the GC-MS analysis (Fig. 4A-G), there was a main peak at 8.5 min in the collected decane phase sample, which corresponded to the reference solution of the standard FOH compound dissolved in decane. Mass spectrometry confirmed that the peak at 8.5 min was FOH (Fig. 4-A). However, the peak was not observed in two-phase culture without introducing BBa_K165025. The formation of FOH from FPP was further confirmed by blocking FPP synthesis. In the GC-MS, the FOH peak was observed in E. coli JM109 (BBa_K165025) culture, whereas no peak was observed with transformed E. coli JM109. It was found that FOH need not only ispA(BBa_K165018) but also MEP(BBa_K165024) in E. coli.We submit new part(BBa_K165025) as producing FOH.

Gas Chromatography/Mass(GC/MS)

菌

Note

In order to be considered for the Best Part Collection award, you must fill out this page.

Did your team make a lot of great parts? Is there a theme that ties all your parts together? Do you have more than 10 parts in this collection? Did you make a CRISPR collection, a MoClo collection, or a collection of awesome pigment parts? Describe your parts collection on this page, so the judges can evaluate you for the Best Part Collection award.