Regulatory

Part:BBa_K2377014:Design

Designed by: Tuoyu Zhou   Group: iGEM17_Lanzhou   (2017-10-23)


zarp, zinc two-component relulatory system


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]


Design Notes

Improved target: BBa_K1755301（K1755024、K1755305、K1755303）

Improved form We replaced a sensor element（Copper-Responsive Two-Component System BBa_K190017）for the Zinc-Responsive Two-Component regulatory system (contains σ54 promoter).

Mechanism of changed sensor The mechanism of Zinc sensor is related to a intrinsic signaling pathway in Ecoli. Which is known that many sensing proteins ZraS immobilized on the membrane of cell. And when Zn2+ show up in periplasmic space, the ions will combine with ZraS and trigger it autophosphorylation. (ZraS*) The phosphorylation signal will transmit from ZraS* to ZraR, then ZraR* come to ZraR(HydG) binding site[1] that will make σ54promoter start downstream transcription. (ZraS* ,ZraR* means activated condition.) 

     Figure 1   the composition of Zinc-Responsive Two-Component regulatory system

So, we substitute Zinc-Responsive Two-Component regulatory system for the copper one in BBa_K1755301. Successfully construct a new heavy metal sensor.

Source

BBa_K1755301（K1755024、K1755305、K1755303） Ｗe got the original parts from our lab, which are designed by 2015 LZU-China iGEM

References

[1] Leonhartsberger S, Huber A, Lottspeich F, et al. The hydH/G Genes from Escherichia coli code for a zinc and lead responsive two-component regulatory system.[J]. Journal of Molecular Biology, 2001, 307(1):93-105. [2] Munson GP1, Lam DL, Outten FW, O'HalloranTV. Identification of acopper-responsive two-component system on the chromosome of Escherichia coli K-12. [3] PedorolluD1,2, The ribB FMN riboswaith from Escherichia coli operates at the transcriptional and translational level and regulates riboflavin biosynthesis. FEBS J. 2015 Aug;282(16):3230-42. doi: 10.1111/febs.13226. Epub 2015 Feb 27.