Difference between revisions of "Part:BBa K3771024"
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<br><b style="font-size:1.3rem">Biology</b><br> | <br><b style="font-size:1.3rem">Biology</b><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. <a href="https://pubmed.ncbi.nlm.nih.gov/16045608/" alt="" target="_blank">[1]</a> 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 <a href="https://pubmed.ncbi.nlm.nih.gov/30516051/" alt="" target="_blank">[ | + | <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. <a href="https://pubmed.ncbi.nlm.nih.gov/16045608/" alt="" target="_blank">[1]</a> 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.<a href="https://pubmed.ncbi.nlm.nih.gov/30516051/" alt="" target="_blank">[2]</a><br> |
+ | <div style="width=100%; display:flex; align-items: center; justify-content: center;"> | ||
+ | <img src="https://2021.igem.org/wiki/images/c/c9/T--NCKU_Tainan--taurine_pathway_1.png" style="width:60%;"> | ||
+ | </div> | ||
+ | <p align="center">Fig. 1. Taurine pathways in <i>E. coli</i></p> | ||
<br><b style="font-size:1.3rem">Usage</b><br> | <br><b style="font-size:1.3rem">Usage</b><br> | ||
<br>We ligased the <i>csad</i> fragment and <i>pspA</i> promoter on the pSU expression vector and transformed it into DH5α to complete construction of the plasmid. | <br>We ligased the <i>csad</i> fragment and <i>pspA</i> promoter on the pSU expression vector and transformed it into DH5α to complete construction of the plasmid. | ||
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<br><b style="font-size:1.3rem">Characterization</b><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="https://2021.igem.org/wiki/images/ | + | <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><b style="font-size:1.3rem">Reference</b><br> | <br><b style="font-size:1.3rem">Reference</b><br> | ||
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<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>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> | ||
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===Usage and Biology=== | ===Usage and Biology=== | ||
Latest revision as of 02:47, 22 October 2021
PpspA-CSAD
Description
This composite part is a component of the IFN-γ sensing system and was 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 fragment and pspA promoter on the pSU expression vector and transformed it into DH5α to complete construction of the plasmid.
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
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 12
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 394
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 179