Composite

Part:BBa_K3771021

Designed by: KAO, YI-CHING   Group: iGEM21_NCKU_Tainan   (2021-10-01)


PpspA-JJU10-6xHis


Description

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

Biology

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 jju-6xHis fragment and pspA promoter on the pSU expression vector and transformed it into DH5α to complete construction of the plasmid. The 6xHis allows for confirmation of JJU expression by western blot using the anti-6xHis antibody.

Characterization

Fig. 2. Colony PCR confirmation of the construction


Reference

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


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Categories
Parameters
None