Difference between revisions of "Part:BBa K5322000:Design"

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===Design Notes===
 
===Design Notes===
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To address the reduced mucus layer thickness and dysbiosis causing inflammation in patients with enteritis, we aim to resolve the dysregulation of the mucosal system from the ground up. A natural high-adhesion protein from marine mussels—mussel foot protein (Mfp)—has garnered our attention, particularly Mfp3, which is one of the proteins with the highest DOPA content among the six mussel proteins. We utilized <i>Escherichia coli</i> BL21(DE3) to establish a production system for Mfp, enhancing adhesion in damaged intestines and regulating gut microbiota. Consequently, we designed the plasmid pET29a-J23119-RBS-Mfp3-T7.
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To ensure that the animal-derived mussel foot protein Mfp3 can be successfully expressed in BL21(DE3), we performed codon optimization on the Mfp3 sequence.
  
 
===Source===
 
===Source===

Revision as of 11:06, 30 September 2024


Constitutive Mfp3 Expression System


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
    Illegal NheI site found at 210
    Illegal NotI site found at 174
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

To ensure that the animal-derived mussel foot protein Mfp3 can be successfully expressed in BL21(DE3), we performed codon optimization on the Mfp3 sequence.

Source

The plasmid pET29a-J23119-RBS-Mfp3-T7 utilizes the pET29a vector, which is commonly employed for robust gene expression in Escherichia coli. The components used in this design, including the J23119 promoter and Mfp3, are derived from the standard biological parts registry of iGEM (BBa_J23119, BBa_K4854000), with the mussel foot protein Mfp3 sourced from natural marine mussels. These parts were selected for their validated functionality and compatibility in synthetic biology applications, facilitating controlled expression within bacterial cells under specific conditions.

References