Part:BBa_K3771039
PpspA-CS-6xHis
Description
This composite part is a component of the IFN-γ sensing system and was used to express the taurine production enzyme, L-cysteine sulfonic acid synthase (CS).
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 CS-his-tag. CS converts o-phospho-l-serine 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 NPO-OmpA* fragment and pspA-CS-his-tag on the pSU expression vector and transformed it into DH5alpha to complete construction of the plasmid. The his-tag allows for confirmation of CS expression by western blot using the anti-6X his-tag 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.xhttps://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.8b05093https://pubmed.ncbi.nlm.nih.gov/30516051/
Sequence and Features
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