Difference between revisions of "Part:BBa K5332003:Design"

 
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===Design Notes===
 
===Design Notes===
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OmpA, which helps the protein to cross the cell membrane, needs to be attached to the N-terminus of the protein to enable CMC to localize on the membrane surface to play an adhesive role after expression in the cell.
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We contributed an innovative foundational component to the Registry platform: the artificial glucan-binding protein CMC, designated as BBa_K5332002. This component ingeniously integrates the mechanism of glucan-binding proteins with the scaffold protein CipC from Clostridium cellulovorans. Additionally, we incorporated the mCherry fluorescent protein sequence and the outer membrane protein A (OmpA) signal peptide, designing proteins with multiple copy numbers to optimize performance. To enhance the persistence of engineered strain FMK in the gut and its response in expressing anti-inflammatory factors, we designed a novel artificial glucan-binding protein CMC by simulating the natural mechanisms of glucan-binding proteins. CMC acts as a "bridge" between gut microbiota and the intestinal surface, effectively attracting and recruiting beneficial gut probiotics. This design not only promotes stable adherence of the engineered strain in the gut but also significantly enhances its efficacy in alleviating intestinal inflammation and modulating the gut microenvironment.
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The copy number of CBMcipc may affect its ability to bind dextran, so different copy numbers of the factor should be designed for adhesion testing, and also the copy number of CBMcipc may have an effect on its cellular distribution, and experiments need to be designed to detect the cellular distribution of the factor.
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Revision as of 07:10, 2 October 2024


CMC (arttificial adhesion protein)


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 470
    Illegal PstI site found at 673
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1444
    Illegal PstI site found at 470
    Illegal PstI site found at 673
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 470
    Illegal PstI site found at 673
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 470
    Illegal PstI site found at 673
    Illegal NgoMIV site found at 381
    Illegal NgoMIV site found at 1806
    Illegal NgoMIV site found at 1810
    Illegal AgeI site found at 295
    Illegal AgeI site found at 1396
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

We contributed an innovative foundational component to the Registry platform: the artificial glucan-binding protein CMC, designated as BBa_K5332002. This component ingeniously integrates the mechanism of glucan-binding proteins with the scaffold protein CipC from Clostridium cellulovorans. Additionally, we incorporated the mCherry fluorescent protein sequence and the outer membrane protein A (OmpA) signal peptide, designing proteins with multiple copy numbers to optimize performance. To enhance the persistence of engineered strain FMK in the gut and its response in expressing anti-inflammatory factors, we designed a novel artificial glucan-binding protein CMC by simulating the natural mechanisms of glucan-binding proteins. CMC acts as a "bridge" between gut microbiota and the intestinal surface, effectively attracting and recruiting beneficial gut probiotics. This design not only promotes stable adherence of the engineered strain in the gut but also significantly enhances its efficacy in alleviating intestinal inflammation and modulating the gut microenvironment.



Source

The core component of CMC, CBMcipc, is derived from the scaffold protein CipC of Clostridium cellulolyticum

References