Difference between revisions of "Part:BBa K3771016"
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In the IFN-γ sensing system, binding of IFN-γ to the OmpA/OprF chimeric protein activates the pspA promoter, producing the enzyme required for the synthesis of taurine.<br>
 
In the IFN-γ sensing system, binding of IFN-γ to the OmpA/OprF chimeric protein activates the pspA promoter, producing the enzyme required for the synthesis of taurine.<br>
  
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<br><b style="font-size:1.3rem">Usage</b>
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<br>The ompA promoter sequence was amplified from E. coli MG1655, digested, and purified. We ligased ompA promoter and the ompA/oprF gene expression vector pSU and transformed it into DH5α to complete construction of the plasmid. Then, the plasmid was purified and transformed into E. coli ompA mutant strain JW0940.
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<br><b style="font-size:1.3rem">Characterization</b>
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<br>PCR<br>
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<div style="width=100%; display:flex; align-items: center; justify-content: center;">
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<img src="https://2021.igem.org/wiki/images/2/20/T--NCKU_Tainan--composite_pOmpA_ompA_oprF.jpg" style="width:35%;">
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<p align="center">圖片描述</p>
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<br>Western blot was performed to confirm the expression of OmpA/OprF.<br>
  
  

Revision as of 13:10, 17 October 2021


PompA-OmpA/OprF


Description


PompA-ompA/oprF is part of the IFN-γ sensing system. ompA promoter regulates the constitutive expression of OmpA/OprF.


Biology
In the IFN-γ sensing system, binding of IFN-γ to the OmpA/OprF chimeric protein activates the pspA promoter, producing the enzyme required for the synthesis of taurine.


Usage


The ompA promoter sequence was amplified from E. coli MG1655, digested, and purified. We ligased ompA promoter and the ompA/oprF gene expression vector pSU and transformed it into DH5α to complete construction of the plasmid. Then, the plasmid was purified and transformed into E. coli ompA mutant strain JW0940.


Characterization


PCR

<img src="T--NCKU_Tainan--composite_pOmpA_ompA_oprF.jpg" style="width:35%;">

圖片描述


Western blot was performed to confirm the expression of OmpA/OprF.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1142
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]