Difference between revisions of "Part:BBa K4055533"
 
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<partinfo>BBa_K4055533 short</partinfo>
 
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Through the combination of three components, we made a blue light photosensitive system. When exposed to blue light,this part can activate the following T7 RNA polymerase.  
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Biofilm structural proteins in E. coli fused with mussel foot  protein(Mfp) analogs bestowed the engineered biofilms with HA mineralization-promoting and  interfacial binding roles. When a blue light-induced strain was used to grow functional biofilms with controlled spatial and biomass density, living mineralized materials with patterning and gradient features could be generated following a benign biomimetic HA mineralization process.  
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In particular,in this circuit, the expression of histidinine kinase YF1 and its homologous regulatory factor FixJ are tightly regulated by LacIq promoter, while the expression repressor cI of phage λ is controlled by pFixK2 promoter. Under blue light irradiation, the kinase activity and subsequent cI expression of YF1 were inhibited, which then activated the λ promoter pR and promoted the expression of CSGA-MFP3S-PEP.
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an Mfp3S protein could initiate HA mineralization and promote interfacial adhesion.CsgA–Mfp fusion proteins comprising a CsgA domain (major protein component of the E. coli biofilm31)  and a C-terminal Mfp serial fusion domain were constructed.
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Moreover, examination of in vivo mineralization by TEM confirmed that CsgA–Mfp3S-pep-expressing biofilms triggered denser mineral formation with more apparent crystalline features after 5 d of min-eralization . These results thus highlight the roles of the Mfp3S-pep fusion protein in promoting HA mineral formation and crystallization.
  
 
===Experiment===
 
===Experiment===
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Experiments in BBa_K4055535 are used to prove the effectiveness of the system
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[[File:T--Yucai_SZ--rs.jpeg|600px|thumb|center|The staining results of a dish of CsgA-Mfp3S-pep bacteria with IPTG (Left) and a blank control dish of amilCP bacteria (Right)]]
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After 9 days of SBF culture, THE MFP-CSGA was scanned by electron microscopy (compared with the sample on day 0).
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[[File:T--Yucai_SZ--2.png|500px|thumb|center|SEM image showing the surface morphology of the biofilm (i.e., unmineralized). Scale bars:3μm]]
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[[File:T--Yucai_SZ--3.png|500px|thumb|center|SEM image showing the surface morphology of the mineralized composite (mineralization 7d). Scale bars: 3μm]]
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[[File:T--Yucai_SZ--kuanghua.jpeg|600px|thumb|center|CsgA-Mfp3S-pep bacteria (Left) and amilCP bacteria (Right) cultured in SBF]]
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here
 
===Usage and Biology===
 
===Usage and Biology===

Latest revision as of 01:02, 22 October 2021


CsgA-Mfp3S-pep

Biofilm structural proteins in E. coli fused with mussel foot protein(Mfp) analogs bestowed the engineered biofilms with HA mineralization-promoting and interfacial binding roles. When a blue light-induced strain was used to grow functional biofilms with controlled spatial and biomass density, living mineralized materials with patterning and gradient features could be generated following a benign biomimetic HA mineralization process.

an Mfp3S protein could initiate HA mineralization and promote interfacial adhesion.CsgA–Mfp fusion proteins comprising a CsgA domain (major protein component of the E. coli biofilm31) and a C-terminal Mfp serial fusion domain were constructed.

Moreover, examination of in vivo mineralization by TEM confirmed that CsgA–Mfp3S-pep-expressing biofilms triggered denser mineral formation with more apparent crystalline features after 5 d of min-eralization . These results thus highlight the roles of the Mfp3S-pep fusion protein in promoting HA mineral formation and crystallization.

Experiment

The staining results of a dish of CsgA-Mfp3S-pep bacteria with IPTG (Left) and a blank control dish of amilCP bacteria (Right)

After 9 days of SBF culture, THE MFP-CSGA was scanned by electron microscopy (compared with the sample on day 0).

SEM image showing the surface morphology of the biofilm (i.e., unmineralized). Scale bars:3μm
SEM image showing the surface morphology of the mineralized composite (mineralization 7d). Scale bars: 3μm
CsgA-Mfp3S-pep bacteria (Left) and amilCP bacteria (Right) cultured in SBF

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]