Difference between revisions of "Part:BBa K4090010"

 
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A kind of fusion protein that helps to form silicon dioxide outside the membrane.
 
A kind of fusion protein that helps to form silicon dioxide outside the membrane.
  
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===Usage and Biology===
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==Data==
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[[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).
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[[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.
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[[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:
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•Both NC Control are darker (NC1 remain as yellow which indicates that it needs MORE NaOH to turn blue, it contains more acids).
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•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).
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[[File:T--SDSZ China--result 1-7parts.jpeg|700px|thumb|center|Fig.4]]
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Fig.1-4 shows the results from TEM testing. Dark proportions represent silicon dioxide produced on the surface of cells.
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[[File:T--SDSZ China--result 1-3parts.png|700px|thumb|center|Fig.5]]
  
 
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<partinfo>BBa_K4090010 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K4090010 SequenceAndFeatures</partinfo>
  
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==References==
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[1] Pamirsky, Igor E., and Kirill S. Golokhvast. "Silaffins of diatoms: from applied biotechnology to biomedicine." Marine drugs 11.9 (2013): 3155-3167.<br>
 +
[2] Park, Jeong Chan, et al. "R5 Peptide-based Biosilicification Using Methyltrimethoxysilane." Biotechnology & Bioprocess Engineering 23.1 (2018).<br>
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[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.<br>
 +
[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.<br>
 +
[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.
  
 
<!-- Uncomment this to enable Functional Parameter display  
 
<!-- Uncomment this to enable Functional Parameter display  

Revision as of 11:11, 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.1-1, 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.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.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.4

Fig.1-4 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 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.