Coding

Part:BBa_K4090001

Designed by: Yiming Dong   Group: iGEM21_SDSZ_China   (2021-10-01)


csgA

Introduction

CsgA is one of the several major components of E.coli biofilm. When it is co-expressed with the gene, it could bring the protein that the gene encodes to the cell surface and expose the protein to the extracellular environment. Sequence and Features


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]

Usage and Viability

To test the function of the CsgA, it was co-expressed with Mfp5, a kind of protein which could induce the precipitation of the calcium carbonate in favorable environment.

Fig.1



After constructing the plasmids by Gibson Assembly, our team used PCR to test whether the plasmids were successfully constructed. Since we omitted a part of the sequence of CsgA at first, we inserted this part of gene later and used the gel electrophoresis to detect this part. The number of bp was exactly the number we expected.

Fig.2



After proving the success of construction, electron diffraction was used to verify the function of the CsgA. The electron-diffaraction diagram below showed the existence of crystal in the sample. Since there was no other crystals in the sample, it could prove the precipitation of the calcium carbonate, which indirectly verified the ability of CsgA in bring the protein to the cell surface.

Fig.3


References

[1] DHAMI N K, REDDY M S, MUKHERJEE A. Biomineralization of calcium carbonate polymorphs by the bacterial strains isolated from calcareous sites[J]. Journal of Microbiology and Biotechnology, 2013, 23(5): 707-714.
[2] OKWADHA G D, LI J. Optimum conditions for microbial carbonate precipitation[J]. Chemosphere, 2010,81(9):1143-1148. [16]LI W, LIU L P, ZHOU P P, et al. Calcite precipitation induced by bacteria and bacterially produced carbonic anhydrase[J]. Current Science, 2011, 100(4): 502 - 508.
[3] ANBU P, KANG C H, SHIN Y J, et al. Formations of calcium carbonate minerals by bacteria and its multiple applications[J]. Springerplus, 2016, 5(1): 1 - 26


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