Difference between revisions of "Part:BBa K3771015"
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<br>When there is no <i>lac</i> repressor, the <i>lacI</i> promoter facilitates the constitutive expression of OmpA/OprF. In the IFN-γ sensing system, binding of IFN-γ to the OmpA/OprF chimeric protein activates the <i>pspA</i> promoter, producing the enzyme required for the synthesis of taurine.  
 
<br>When there is no <i>lac</i> repressor, the <i>lacI</i> promoter facilitates the constitutive expression of OmpA/OprF. In the IFN-γ sensing system, binding of IFN-γ to the OmpA/OprF chimeric protein activates the <i>pspA</i> promoter, producing the enzyme required for the synthesis of taurine.  
 
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  <br><b style="font-size:1.3rem">Usage</b>
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  <br>We ligased <i>ompA/oprF</i> gene and the <i>P<sub>lacI</sub></i> expression vector pSU and transformed it into DH5α to complete construction of the plasmid. Then, the plasmid was purified and transformed into <i>E. coli ompA</i> mutant strain JW0940.<br>
  
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<br><b style="font-size:1.3rem">Characterization</b> 
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<img src="https://2021.igem.org/wiki/images/e/e6/T--NCKU_Tainan--composite_pLacI_ompA_oprF_.jpg
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<p align="center">PCR</p>
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<br>Western blot was performed to confirm the expression of OmpA/OprF
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Revision as of 11:39, 17 October 2021


PlacI-OmpA/OprF

Description

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

Biology

When there is no lac repressor, the lacI promoter facilitates the constitutive expression of OmpA/OprF. 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
We ligased ompA/oprF gene and the PlacI 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


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 1039
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]