Composite

Part:BBa_K3771045

Designed by: KAO, YI-CHING   Group: iGEM21_NCKU_Tainan   (2021-10-01)
Revision as of 16:01, 17 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].


Usage


We ligased the PompA-ompA/oprF fragment and PpspA-CDO1 on the pSU expression vector and transformed it into DH5α to complete construction of the plasmid.


Characterization


Double digestion results are shown in Fig. 2.


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/
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