Difference between revisions of "Part:BBa K3110054"
 
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<partinfo>BBa_K3110054 short</partinfo>
 
<partinfo>BBa_K3110054 short</partinfo>
  
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YebF is a 13kDa protein that is known to be secreted into the extracellular medium by E. coli strains used in the lab. This particular protein was used with IL-12 in our design, in order to mediate the secretion of the cytokine from the bacteria. The part was used with constitutive promoter (J23100) but the results were inconclusive. Western blots were done with both the cell pellet and concentrated supernatant but the protein was not detected.Possible errors could be because of SOEing errors in the promoter and RBS regions.
 
  
 
The original YebF part can be found here: BBa_K1659003  
 
The original YebF part can be found here: BBa_K1659003  
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===Usage and Biology===
 
  
 
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<partinfo>BBa_K3110054 parameters</partinfo>
 
<partinfo>BBa_K3110054 parameters</partinfo>
 
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<h1>Usage and Biology</h1>
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YebF is a 13 kDa protein that is known to be secreted into the extracellular medium by <i>E. coli</i> strains used in the lab. This particular protein was conjugated with IL-12 in our design, in order to mediate the secretion of the cytokine from the bacteria. The part was used with a constitutive promoter (J23100) but the results were inconclusive.SDS-PAGE was done with both the cell pellet and concentrated supernatant of our test sample and a control (untransformed BL21) but no visible band difference was detected. We suspected this could be because of SOEing errors in the promoter and RBS regions.
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<h1>Characterization</h1>
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This part was used as a control for O1PO2-StrongRBS-YebF-IL12-FLAG ([[Part:BBa_K3110051]]) to check the difference between YebF-IL12 produced by the inducible promoter and constitutive Promoter (without the operator regions).
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The results for this construct can be found with the Result of the sample construct ([[Part:BBa_K3110051]])

Latest revision as of 02:55, 22 October 2019


J23117-YebF IL12-FLAG


The original YebF part can be found here: BBa_K1659003

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Usage and Biology

YebF is a 13 kDa protein that is known to be secreted into the extracellular medium by E. coli strains used in the lab. This particular protein was conjugated with IL-12 in our design, in order to mediate the secretion of the cytokine from the bacteria. The part was used with a constitutive promoter (J23100) but the results were inconclusive.SDS-PAGE was done with both the cell pellet and concentrated supernatant of our test sample and a control (untransformed BL21) but no visible band difference was detected. We suspected this could be because of SOEing errors in the promoter and RBS regions.

Characterization

This part was used as a control for O1PO2-StrongRBS-YebF-IL12-FLAG (Part:BBa_K3110051) to check the difference between YebF-IL12 produced by the inducible promoter and constitutive Promoter (without the operator regions). The results for this construct can be found with the Result of the sample construct (Part:BBa_K3110051)