Coding

Part:BBa_K4090000

Designed by: Yiming Dong   Group: iGEM21_SDSZ_China   (2021-10-01)
Revision as of 10:17, 17 October 2021 by Yi Rao (Talk | contribs)

Ice Nucleation Protein(INP)

ice nucleation protein(INP) is a kind of transmembrane protein, which can help proteins to play their role outside the membrane.

Data

Fig.1-1

According to Fig.1-1, 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.1-2

Fig.1-2 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.1-3

Fig.1-3 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.1-4

Fig.1-4 shows the results from TEM testing. Dark proportions represent silicon dioxide produced on the surface of cells.

Fig.1-5



Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 321
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 63
    Illegal NgoMIV site found at 396
    Illegal AgeI site found at 498
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 229

References

[1]ZHU T, PAULO C, MERROUN, M L, et al. Potential application of biomineralization by Synechococcus PCC8806 for concrete restoration[J]. Sedimentology, 2013,61(1):1-21
[2]Lechner, Carolin C., and Christian FW Becker. "A sequence‐function analysis of the silica precipitating silaffin R5 peptide." Journal of Peptide Science 20.2 (2014): 152-158.
[3]Poulsen, Nicole, and Nils Kroger. "Silica morphogenesis by alternative processing of silaffins in the diatom Thalassiosira pseudonana." Journal of Biological Chemistry 279.41 (2004): 42993-42999.


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