Difference between revisions of "Part:BBa K3771035"

 
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<partinfo>BBa_K3771035 short</partinfo>
 
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<br><b style="font-size:1.3rem">Description</b>
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<br>This composite part is a component of the IFN-γ sensing system and was used to express the taurine production enzyme, CoaBC.
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<br><b style="font-size:1.3rem">Biology</b>
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<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 CoaBC. CoaBC converts L-cystate into taurine in the taurine synthesis L-cystate pathway.
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<br><b style="font-size:1.3rem">Usage</b>
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<br>We ligased the NPO-OmpA* fragment and pspA-CoaBC-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 CoaBC expression by western blot using the anti-6X his-tag antibody.
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<br><b style="font-size:1.3rem">Characterization</b>
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<br>Double digestion results are shown in Fig. 2.
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<div style="width=100%; display:flex; align-items: center; justify-content: center;">
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<img src="圖片網址" style="width:35%;">
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<p align="center">圖片描述</p>
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<br><b style="font-size:1.3rem">References</b>
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<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
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<a href="https://pubmed.ncbi.nlm.nih.gov/16045608/" alt="" target="_blank">https://pubmed.ncbi.nlm.nih.gov/16045608/</a>
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<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Revision as of 20:08, 17 October 2021


PpspA-CoaBC-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, CoaBC.


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 CoaBC. CoaBC converts L-cystate into taurine in the taurine synthesis L-cystate pathway.


Usage


We ligased the NPO-OmpA* fragment and pspA-CoaBC-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 CoaBC expression by western blot using the anti-6X his-tag antibody.


Characterization


Double digestion results are shown in Fig. 2.

<img src="圖片網址" style="width:35%;">

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 <a href="https://pubmed.ncbi.nlm.nih.gov/16045608/" alt="" target="_blank">https://pubmed.ncbi.nlm.nih.gov/16045608/</a>

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 2533
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 727
  • 1000
    COMPATIBLE WITH RFC[1000]