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<br>The LacI promoter facilitates the constitutive expression of OmpA/OprF. Binding of IFN-γ to the OmpA/OprF chimeric protein induces the response of the phage shock protein (Psp) system, a highly conserved stress response system in enterobacteria[1]. Signal transduction from the outer membrane to the inner membrane activates the <i>pspA</i> promoter, initiating expression of JJU. JJU converts L-cysteine into L-cystate in the taurine synthesis L-cystate pathway.
 
<br>The LacI promoter facilitates the constitutive expression of OmpA/OprF. Binding of IFN-γ to the OmpA/OprF chimeric protein induces the response of the phage shock protein (Psp) system, a highly conserved stress response system in enterobacteria[1]. Signal transduction from the outer membrane to the inner membrane activates the <i>pspA</i> promoter, initiating expression of JJU. JJU converts L-cysteine into L-cystate in the taurine synthesis L-cystate pathway.
 
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<br>Binding of IFN-γ to the OmpA/OprF chimeric protein induces the response of the phage shock protein (Psp) system, a highly conserved stress response system in enterobacteria. <a href="https://pubmed.ncbi.nlm.nih.gov/16045608/" alt="" target="_blank">[1]</a> Signal transduction from the outer membrane to the inner membrane activates the pspA promoter, initiating expression of JJU. JJU catalyzes the decarboxylation of L-Cysteine sulfinic acid into hypotaurine, which is spontaneously oxidized to taurine <a href="https://pubmed.ncbi.nlm.nih.gov/30516051/" alt="" target="_blank">[2]</a>. <br>
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<img src="https://2021.igem.org/wiki/images/c/c9/T--NCKU_Tainan--taurine_pathway_1.png" style="width:60%;">
 
<img src="https://2021.igem.org/wiki/images/c/c9/T--NCKU_Tainan--taurine_pathway_1.png" style="width:60%;">

Revision as of 21:23, 19 October 2021


PpspA-JJU-PlacI-OmpA/OprF
Description

This composite part is a component of the IFN-γ sensing system and is used to express the taurine production enzyme, JJU.

Biology

The LacI promoter facilitates the constitutive expression of OmpA/OprF. Binding of IFN-γ to the OmpA/OprF chimeric protein induces the response of the phage shock protein (Psp) system, a highly conserved stress response system in enterobacteria[1]. Signal transduction from the outer membrane to the inner membrane activates the pspA promoter, initiating expression of JJU. JJU converts L-cysteine into L-cystate in the taurine synthesis L-cystate pathway.

Fig.1 Taurine pathways in E. coli


Usage

We ligased the PlacI-ompA/oprF fragment and PpspA-jju on the pSU expression vector and transformed it into DH5α to complete construction of the plasmid.

Characterization

Fig. 2. Double digestion check of PpspA-jju

Fig. 3. Colony PCR confirmation of the construction


References

1. Darwin AJ. The phage-shock-protein response. Molecular Microbiology. 2005;57(3):621-628. doi:10.1111/j.1365-2958.2005.04694.x https://pubmed.ncbi.nlm.nih.gov/16045608/

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 12
    Illegal BamHI site found at 2364
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]