Composite

Part:BBa_K3771018

Designed by: KAO, YI-CHING   Group: iGEM21_NCKU_Tainan   (2021-10-01)
Revision as of 07:07, 17 October 2021 by Evelynnn (Talk | contribs)


PpspA-OFP-PompA-OmpA/OprF
Description

This composite part is part of a plasmid used in the IFN-γ induction assay to test the function of the IFN-γ sensing system. It consists of the ompA/oprF gene regulated by the ompA promoter (PompA-ompA/oprF) and the orange fluorescent protein (OFP) gene regulated by the pspA promoter (PpspA-ofp).

Biology

Figure. 1 Structure of OmpA/OprF chimeric protein


To allow E. coli to detect IFN-γ in the human gut, we constructed an OmpA/OprF chimeric protein designed by Aurand and March[1]. The OmpA/OprF chimeric protein consists of OmpA protein from E. coli and a smaller section of OprF protein from P. aeruginosa.

Figure 2. Mechanism of IFN-γ sensing system


The ompA promoter facilitates the constitutive expression of chimeric OmpA/OprF. Binding of IFN-γ to chimeric OmpA/OprF induces the response of the phage shock protein (Psp) system[1], a highly conserved stress response system in enterobacteria[2]. Signal transduction from the outer membrane to the inner membrane activates the pspA promoter, initiating expression of OFP.
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 2158
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
None