Part:BBa_K4090002

Mfp5

Introduction

Mfp5 is found in mussel foot. It helps mussel tightly attach to the rock by inducing the precipitation of the hydroxyapatite. It could also induce the precipitation in vitro in the 1.5×SBF solution. Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 73
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]

Usage and Viability

Since the Mfp5 is functional only when it is exposed to the favorable environment,it was co-expressed with CsgA, a kind of biofilm protein which could bring the protein to the cell surface by co-expression.

Fig.2

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.

Fig.1

After proving the success of construction, electron diffraction was used to verify the function of the Mfp5 in inducing the precipitation. The electron-diffraction 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 hydroxyapatite, which indirectly verified the ability of Mfp5 in inducing the precipitation of hydroxyapatite.

Fig.3

References

[1] 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.
[2] 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.
[3] 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.