Composite

Part:BBa_K4767007

Designed by: Fangjun Liu   Group: iGEM23_CUG-China   (2023-10-04)


Pars-RBS-arsR-RBS-mtrC-TT

Uses a factor Pars(BBa_K4767001) ,strong RBS(BBa_J34801), DNA binding transcriptional repressor arsR(BBa_J15101), c-type cytochromes mtrC(BBa_K3102020) and TT(BBa_B0015). This composite part is used in the iron reduction experiment and MFC.

Usage and Biology

This part is a arsenic reporter with the reporter gene mtrC. MtrC is a c-type cytochromes from Shewanella oneidensis. It plays an important role in its extracellular electron transfer. To verify this part can respond to arsenic, we transfer this part into a S. oneidensis strain with the mtrC gene mutant. Then we cultured the strain with a arsenic concentration gradient. When arsenic is absent, the transcription regulator ArsR binds to the ArsR-binding site (ABS) within the ars promoter and blocks transcription. Once arsenic is present, it binds to ArsR and activate the transcription of the mtrC genes. As the result showed, strains with higher arsenic has a faster iron reduction rate, indicating the more expression of mtrC.

img-1170.png
Fig. Iron reduction curves with different arsenic concentrations

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 270
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 751

References

Xiaoqiang Jia, Bu Rongrong, Zhao Tingting, et al. Sensitive and Specific Whole-Cell Biosensor for Arsenic Detection[J]. Applied and environmental microbiology, 2019, 85(11): 1.

[edit]
Categories
//classic/reporter
Parameters
biologyS. oneidensis & Escherichia coli