Difference between revisions of "Part:BBa K3771043"

Revision as of 22:03, 19 October 2021 (view source) Evelynnn (Talk | contribs) ← Older edit		Revision as of 22:06, 19 October 2021 (view source) Evelynnn (Talk | contribs) Newer edit →
Line 25:		Line 25:
	<br>		<br>
−	<br><b style="font-size:1.3rem">Characterization</b>
−	<br>
−	<br>Double digestion results are shown in Figure 2.
−	<br>
−
−	<div style="width=100%; display:flex; align-items: center; justify-content: center;">
−	<img src="https://2021.igem.org/wiki/images/d/d7/T--NCKU_Tainan--pspAcsadv.jpg" style="width:40%;">
−	</div>
−	<p align="center">Fig. 2. Double digestion check of <i>P<sub>pspA</sub>-csad</i></p>
−	<div style="width=100%; display:flex; align-items: center; justify-content: center;">
−	<img src="https://2021.igem.org/wiki/images/2/22/T--NCKU_Tainan--colony_pspAcsadompA.jpg
−	" style="width:40%;">
−	</div>
−	<p align="center">Fig. 3. Colony PCR confirmation of the construction</p>
	<br><b style="font-size:1.3rem">References</b>		<br><b style="font-size:1.3rem">References</b>
	<br>		<br>

---

Revision as of 22:06, 19 October 2021

PpspA-CS-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, 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 P_ompA-ompA/oprF fragment and P_pspA-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