Difference between revisions of "Part:BBa K3771039"
Marvinang02 (Talk | contribs) |
|||
| Line 4: | Line 4: | ||
<html> | <html> | ||
| − | <br><b style="font-size:1.3rem">Description</b> | + | <br><b style="font-size:1.3rem">Description</b><br> |
| − | <br> | + | <br>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). <br> |
| − | + | <br><b style="font-size:1.3rem">Biology</b><br> | |
| − | <br>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). | + | <br>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].<div style="width=100%; display:flex; align-items: center; justify-content: center;"> |
| − | <br> | + | <img src="https://2021.igem.org/wiki/images/c/c9/T--NCKU_Tainan--taurine_pathway_1.png" style="width:60%;"> |
| − | + | </div> | |
| − | <br><b style="font-size:1.3rem">Biology</b> | + | <p align="center">Fig.1 Taurine pathways in <i>E. coli</i></p> |
| − | <br> | + | <br><b style="font-size:1.3rem">Usage</b><br> |
| − | + | <br>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.<br> | |
| − | <br>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] | + | <br><b style="font-size:1.3rem">Characterization</b><br> |
| − | < | + | |
| − | + | ||
| − | <br><b style="font-size:1.3rem">Usage</b> | + | |
| − | <br> | + | |
| − | + | ||
| − | <br>We ligased the CS-his-tag | + | |
| − | <br> | + | |
| − | + | ||
| − | <br><b style="font-size:1.3rem">Characterization</b> | + | |
| − | <br> | + | |
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
<div style="width=100%; display:flex; align-items: center; justify-content: center;"> | <div style="width=100%; display:flex; align-items: center; justify-content: center;"> | ||
| − | <img src=" | + | <img src="https://2021.igem.org/wiki/images/4/46/T--NCKU_Tainan--colony_csad.jpg" style="width:40%;"> |
</div> | </div> | ||
| − | <p align="center"> | + | <p align="center">Fig. 2. Colony PCR confirmation of the construction</p> |
| − | + | ||
| − | + | ||
| − | + | ||
| − | <br>1. Darwin AJ. The phage-shock-protein response. | + | <br><b style="font-size:1.3rem">Reference</b><br> |
| − | <a href="https://pubmed.ncbi.nlm.nih.gov/16045608/" alt="" target="_blank">https://pubmed.ncbi.nlm.nih.gov/16045608/</a> | + | <br>1. Darwin AJ. The phage-shock-protein response. Molecular Microbiology. 2005;57(3):621-628. doi:10.1111/j.1365-2958.2005.04694.x<a href="https://pubmed.ncbi.nlm.nih.gov/16045608/ |
| − | <br> | + | " alt="" target="_blank">https://pubmed.ncbi.nlm.nih.gov/16045608/ |
| − | <br>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. | + | </a><br> |
| − | <a href="https://pubmed.ncbi.nlm.nih.gov/30516051/" alt="" target="_blank">https://pubmed.ncbi.nlm.nih.gov/30516051/</a> | + | <br>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<a href="https://pubmed.ncbi.nlm.nih.gov/30516051/" alt="" target="_blank">https://pubmed.ncbi.nlm.nih.gov/30516051/</a><br> |
| − | <br | + | |
| − | + | ||
<!-- Add more about the biology of this part here | <!-- Add more about the biology of this part here | ||
===Usage and Biology=== | ===Usage and Biology=== | ||
Revision as of 21:44, 19 October 2021
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