Composite

Part:BBa_K4090007

Designed by: Yiming Dong   Group: iGEM21_SDSZ_China   (2021-10-01)
Revision as of 19:02, 21 October 2021 by Vague (Talk | contribs)


csgA-Mfp5-Mfp5

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. Mfp5 is found in mussel foot. It could induce the precipitation in vitro in the 1.5×SBF solution. According to previous experiments, the expression level of one Mfp5 gene may be insufficient to induce notable precipitation, two Mfp5 genes are incorporated in this part. This biobrick could induce the precipitation of hydroxyapatite.

Usage and Viability

We designed the part as below.

Fig.1



After constructing the plasmids by Gibson Assembly, our team used PCR to test whether the plamids 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, proving that the construction was successful.

Since the lack of the time, we failed to carried out the experimnets to prove the function of this composite part.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 371
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 371
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 556
    Illegal BamHI site found at 784
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 371
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 371
  • 1000
    COMPATIBLE WITH RFC[1000]

Reference

[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.
[4] Hale, L. V., Ma, Y. F. & Santerre, R. F. Semi-quantitative fuorescence analysis of calcein binding as a measurement of in vitro mineralization. Calcif. Tissue Int. 67, 80-84(2000). Ramachandran S K.
[5] Dick J, Windt W D, Graef B D, et al. Bio-deposition of a calcium carbonate layer on degraded limestone by Bacillus species[J]. Biodegradation, 2016(4): 357-367.
[6] Qian C X, Chen H C, Ren L F, et al. Self-healing of early age cracks in cement-based materials by mineralization of carbonic anhydrase microorganism[J]. Frontiers in Microbiology, 2015(6): 1-9.


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