Difference between revisions of "Part:BBa K3771044"

 
Line 3: Line 3:
 
<partinfo>BBa_K3771044 short</partinfo>
 
<partinfo>BBa_K3771044 short</partinfo>
  
−
to be edited
+
<html>
 +
<br><b style="font-size:1.3rem">Description</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>
 +
 +
<br><b style="font-size:1.3rem">Biology</b>
 +
<br>
 +
 +
<br>The <i>ompA</i> 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 <i>pspA</i> 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].
 +
<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 DH5α 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><b style="font-size:1.3rem">Characterization</b>
 +
<br>
 +
 +
<br>Double digestion results are shown in Figure 1.
 +
<br>
 +
 +
<div style="width=100%; display:flex; align-items: center; justify-content: center;">
 +
<img src="圖片網址" style="width:35%;">
 +
</div>
 +
<p align="center">圖片描述</p>
 +
 +
<br><b style="font-size:1.3rem">References</b>
 +
<br>
 +
 +
<br>1. Darwin AJ. The phage-shock-protein response. <i>Molecular Microbiology</i>. 2005;57(3):621-628. doi:10.1111/j.1365-2958.2005.04694.x
 +
<a href="https://pubmed.ncbi.nlm.nih.gov/16045608/" alt="" target="_blank">https://pubmed.ncbi.nlm.nih.gov/16045608/</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. <i>Journal of Agricultural and Food Chemistry</i>. 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>
 +
</html>
 
<!-- 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 20:44, 17 October 2021


PpspA-CS-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, 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].

Usage

We ligased the NPO-OmpA* fragment and pspA-CS-his-tag on the pSU expression vector and transformed it into DH5α 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

Double digestion results are shown in Figure 1.

圖片描述


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


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 12
    Illegal BamHI site found at 1915
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 591
  • 1000
    COMPATIBLE WITH RFC[1000]