Composite

Part:BBa_K3771045

Designed by: KAO, YI-CHING   Group: iGEM21_NCKU_Tainan   (2021-10-01)
Revision as of 22:11, 19 October 2021 by Evelynnn (Talk | contribs)


PpspA-CDO1-pOmpA-OmpA/OprF


Description

This composite part is a component of the IFN-γ sensing system and was used to express the taurine production enzyme, CDO1.

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 CDO1. CDO1 converts L-cysteine into L-cysteine sulfonic acid in the taurine synthesis L-cysteine sulfinic acid pathway[2].

Fig.1 Taurine pathways in E. coli


Usage

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

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/

2. Joo Y-C, Ko YJ, You SK, et al. Creating a New Pathway in Corynebacterium glutamicum for the Production of Taurine as a Food Additive. Journal of Agricultural and Food Chemistry. 2018;66(51):13454-13463. doi:10.1021/acs.jafc.8b05093 https://pubmed.ncbi.nlm.nih.gov/30516051/
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 1897
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 591
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
None