Difference between revisions of "Part:BBa K3771033"
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<br>The ompA 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. [3] Signal transduction from the outer membrane to the inner membrane activates the <i>pspA</i> promoter, initiating expression of JJU10. JJU10 converts L-cysteine into L-cystate in the taurine synthesis L-cystate pathway.
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<br>The ompA 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 JJU10. JJU10 converts L-cysteine into L-cystate in the taurine synthesis L-cystate pathway.
  
 
<br><b style="font-size:1.3rem">Usage</b>
 
<br><b style="font-size:1.3rem">Usage</b>

Revision as of 19:56, 17 October 2021


PpspA-JJU-6xHis-PompA-OmpA/OprF


Description

This composite part is a component of the IFN-γ sensing system and is used to express the taurine production enzyme, JJU10.

Biology

The ompA 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 JJU10. JJU10 converts L-cysteine into L-cystate in the taurine synthesis L-cystate pathway.
Usage

We ligased the NPO-OmpA* fragment and pspA-JJU10-his-tag on the pSU expression vector and transformed it into DH5alpha to complete construction of the plasmid. The his-tag allows for confirmation of JJU10 expression by western blot using the anti-6X his-tag antibody.

Characterization

Double digestion results are shown in Figure 1.

圖片描述


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 2485
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]