Difference between revisions of "Part:BBa K4090010"
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==Data==
 
==Data==
 
[[File:T--SDSZ_China--result_1-2parts.png|700px|thumb|center|Fig.1]]
 
[[File:T--SDSZ_China--result_1-2parts.png|700px|thumb|center|Fig.1]]
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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).
+
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).
 
[[File:T--SDSZ China--result 1-4parts.png|700px|thumb|center|Fig.2]]
 
[[File:T--SDSZ China--result 1-4parts.png|700px|thumb|center|Fig.2]]
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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.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.
 
[[File:T--SDSZ China--result 1-6parts.png|700px|thumb|center|Fig.3]]
 
[[File:T--SDSZ China--result 1-6parts.png|700px|thumb|center|Fig.3]]
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Fig.1-3 represents qualitative OD values, showing that there is sufficient amount of silicatein produced, since:  
+
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 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).
 
•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).
 
[[File:T--SDSZ China--result 1-7parts.jpeg|700px|thumb|center|Fig.4]]
 
[[File:T--SDSZ China--result 1-7parts.jpeg|700px|thumb|center|Fig.4]]
−
Fig.1-4 shows the results from TEM testing. Dark proportions represent silicon dioxide produced on the surface of cells.
+
Fig.4 shows the results from TEM testing. Dark proportions represent silicon dioxide produced on the surface of cells.
−
[[File:T--SDSZ China--result 1-3parts.png|700px|thumb|center|Fig.5]]
+
[[File:T--SDSZ China--result inpsilicatein.png|700px|thumb|center|Fig.5]]
 +
[[File:T--SDSZ China--result 1-3parts.png|700px|thumb|center|Fig.6]]
  
 
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Revision as of 11:13, 17 October 2021


INP-Silicatein

A kind of fusion protein that helps to form silicon dioxide outside the membrane.


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
Fig.6

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1392
  • 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] Pamirsky, Igor E., and Kirill S. Golokhvast. "Silaffins of diatoms: from applied biotechnology to biomedicine." Marine drugs 11.9 (2013): 3155-3167.
[2] Park, Jeong Chan, et al. "R5 Peptide-based Biosilicification Using Methyltrimethoxysilane." Biotechnology & Bioprocess Engineering 23.1 (2018).
[3] Müller, Werner EG, et al. "Bioencapsulation of living bacteria (Escherichia coli) with poly (silicate) after transformation with silicatein-α gene." Biomaterials 29.7 (2008): 771-779.
[4] 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.
[5] 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.