Difference between revisions of "Part:BBa K3771025"
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<br><b style="font-size:1.3rem">Characterization</b> | <br><b style="font-size:1.3rem">Characterization</b> | ||
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<p align="center">Fig. 2. Colony PCR confirmation of the construction</p> | <p align="center">Fig. 2. Colony PCR confirmation of the construction</p> | ||
Revision as of 20:38, 19 October 2021
PpspA-CSAD-6xHis
Description
This composite part is a component of the IFN-γ sensing system and is used to express the taurine production enzyme, CSAD.
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 CSAD. CSAD catalyzes the decarboxylation of L-Cysteine sulfinic acid into hypotaurine, which is spontaneously oxidized to taurine [2].
Fig.1 Taurine pathways in E. coli
Usage
We ligased the csad-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 CSAD 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/
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