Difference between revisions of "Part:BBa K3771046"

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−	<br>We ligased the <i>P<sub>ompA</sub>-ompA/oprF</i> fragment and <i>P<sub>pspA</sub>-cdo1</i> on the pSU expression vector and transformed it into DH5α to complete construction of the plasmid.	+	<br>We ligased the <i>P<sub>ompA</sub>-ompA/oprF</i> fragment and <i>P<sub>pspA</sub>-cdo1-6xHis</i> on the pSU expression vector and transformed it into DH5α to complete construction of the plasmid.
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Revision as of 22:14, 19 October 2021

PpspA-CDO1-6xHis-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 CDO1-6xHis. 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 P_ompA-ompA/oprF fragment and P_pspA-cdo1-6xHis 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