Composite
Part:BBa_K3771018
Designed by: KAO, YI-CHING Group: iGEM21_NCKU_Tainan (2021-10-01)
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:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 12
Illegal BamHI site found at 2158 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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Categories
Parameters
None |