Difference between revisions of "Part:BBa K5127013:Design"

 
 
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===Source===
 
===Source===
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The leucine-responsive regulatory protein Lrp was constitutively expressed as driven by J23101. Then, Lrp activates the expression of cI repressor in the presence of butyrate, and the cI protein inhibits promoter pLam activity (Figure 1). The pCI-Lam, PpchA, protein CI, and GFP were synthesized by Genscript. The Lrp protein was supported by NMU-China.  
 
The leucine-responsive regulatory protein Lrp was constitutively expressed as driven by J23101. Then, Lrp activates the expression of cI repressor in the presence of butyrate, and the cI protein inhibits promoter pLam activity (Figure 1). The pCI-Lam, PpchA, protein CI, and GFP were synthesized by Genscript. The Lrp protein was supported by NMU-China.  
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===References===
 
===References===
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Kineret Serebrinsky-Duek, Barra, M., Danino, T., & Garrido, D. (2023). Engineered Bacteria for Short-Chain-Fatty-Acid-Repressed Expression of Biotherapeutic Molecules. Microbiology Spectrum, 11(2). https://doi.org/10.1128/spectrum.00049-23

Latest revision as of 07:58, 2 October 2024


Butyrate sensor with NOT gate (pPcha-ci-pLam-GFP)


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 970
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 239
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Initially, our plasmid's RBS component, synthesized by Genscript, had an unwanted point mutation. But we did characterization to see whether it could still function well (Figure 2).


Source

The leucine-responsive regulatory protein Lrp was constitutively expressed as driven by J23101. Then, Lrp activates the expression of cI repressor in the presence of butyrate, and the cI protein inhibits promoter pLam activity (Figure 1). The pCI-Lam, PpchA, protein CI, and GFP were synthesized by Genscript. The Lrp protein was supported by NMU-China.


References

Kineret Serebrinsky-Duek, Barra, M., Danino, T., & Garrido, D. (2023). Engineered Bacteria for Short-Chain-Fatty-Acid-Repressed Expression of Biotherapeutic Molecules. Microbiology Spectrum, 11(2). https://doi.org/10.1128/spectrum.00049-23