Part:BBa_K4090004

Silicatein

Silicatein is a kind of mineralized protein that helps to form silicon dioxide

Data

Fig.1

According to Fig.1, all the fragments designed have been successfully made, including S1&S2, which are Silicatein combined with INP, and P1&P2, which are S010_pET28b-csgA-linker-mfp5-7His(pET).

Fig.2

Fig.2 represents the results of gel electrophoresis to test the fragment after Gibson Assembly. The length is about 600bp, which, together with the results from sequencing, indicates a positive outcome.

Fig.3

Fig.3 represents qualitative OD values, showing that there is sufficient amount of silicatein produced, since: •Both NC Control are darker (NC1 remain as yellow which indicates that it needs MORE NaOH to turn blue, it contains more acids). •Both silicatein mixture (with / without homogenization) have higher OD810nm, meaning that there are more bacteria inside the test tube (the solution is thicker / not as clear as NCs).

Fig.4

Fig.4 shows the results from TEM testing. Dark proportions represent silicon dioxide produced on the surface of cells.

Fig.5

Sequence and Features

Assembly Compatibility:

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

  Illegal NheI site found at 664
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]

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.