Difference between revisions of "Part:BBa K3777018"
 
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__NOTOC__
 
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<partinfo>BBa_K3777013 short</partinfo>
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<partinfo>BBa_K3777018 short</partinfo>
  
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Basic biosensor device for tetracycline detection.
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Basic biosensor device for Chlortetracycline detection.
  
 
<!-- Add more about the biology of this part here-->
 
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<b><font size="3">Usage and Biology</font></b>
 
<b><font size="3">Usage and Biology</font></b>
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<br>The genetic circuit was composed of a coding sequence of  tetracycline repressor which was inserted into an expression vectors with a consitive promoter(BBa_J23114) and RBS(BBa_K3777030), as well as 3WJdB(BBa_K3777000) under the control of T7 promoter (BBa_K3777006).  The terminator we used were BBa_B0010 and BBa_M36305.(Fig 1)
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<br>The genetic circuit was composed of a coding sequence of  Chlortetracycline repressor tetM genes which was inserted into an expression vectors with a consitive promoter(BBa_J23114) and RBS(BBa_K3777030), as well as 3WJdB(BBa_K3777000) under the control of T7 promoter (BBa_K3777006).  The terminator we used were BBa_B0010 and BBa_M36305.(Fig 1)
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<br>When tet was absent, TetR would bind to the inducible promoter（PI）and prevent RNA polymerase from initiating transcription, thus repressing the expression of reporter gene. If tet was present, TetR would no longer able to bind to the promoter, resulting in the expression of reporter gene.
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<br>When CTE was absent, TetR would bind to the inducible promoter（PI）and prevent RNA polymerase from initiating transcription, thus repressing the expression of reporter gene. If tet was present, TetR would no longer able to bind to the promoter, resulting in the expression of reporter gene.
 
<br>We expressed this circuit in the <i>E. coli </i> BL21(DE3) cells for tetracycline detection. Thus we could roughly deduce the concentration of the antibiotics in the sample according to the fluorescence intensity.  
 
<br>We expressed this circuit in the <i>E. coli </i> BL21(DE3) cells for tetracycline detection. Thus we could roughly deduce the concentration of the antibiotics in the sample according to the fluorescence intensity.  
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https://static.igem.org/mediawiki/parts/thumb/3/3d/Tetr-tetO-3WJdB.PNG/799px-Tetr-tetO-3WJdB.PNG  
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https://static.igem.org/mediawiki/parts/thumb/c/c9/CtcS-tetm-T7%28ctcO%29-3WJDB.PNG/798px-CtcS-tetm-T7%28ctcO%29-3WJDB.PNG  
 
<br>                          Fig.1  Schematic overview of the genetic circuit.
 
<br>                          Fig.1  Schematic overview of the genetic circuit.
 
<br><b><font size="3">Results</font></b>
 
<br><b><font size="3">Results</font></b>
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
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<partinfo>BBa_K3777013 SequenceAndFeatures</partinfo>
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<partinfo>BBa_K3777018 SequenceAndFeatures</partinfo>
  
  
 
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===Functional Parameters===
 
===Functional Parameters===
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<partinfo>BBa_K3777013 parameters</partinfo>
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<partinfo>BBa_K3777018 parameters</partinfo>
 
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Latest revision as of 17:38, 21 October 2021

ctcS-tetM-T7(ctcO)-3WJdB

Basic biosensor device for Chlortetracycline detection.

Usage and Biology
The genetic circuit was composed of a coding sequence of Chlortetracycline repressor tetM genes which was inserted into an expression vectors with a consitive promoter(BBa_J23114) and RBS(BBa_K3777030), as well as 3WJdB(BBa_K3777000) under the control of T7 promoter (BBa_K3777006). The terminator we used were BBa_B0010 and BBa_M36305.(Fig 1)
When CTE was absent, TetR would bind to the inducible promoter（PI）and prevent RNA polymerase from initiating transcription, thus repressing the expression of reporter gene. If tet was present, TetR would no longer able to bind to the promoter, resulting in the expression of reporter gene.
We expressed this circuit in the E. coli BL21(DE3) cells for tetracycline detection. Thus we could roughly deduce the concentration of the antibiotics in the sample according to the fluorescence intensity. 798px-CtcS-tetm-T7%28ctcO%29-3WJDB.PNG
Fig.1 Schematic overview of the genetic circuit.
Results
To verify the functionality of the biosensor, we performed a plate-reader experiment and measured optical density and fluorescence intensity every hour. We observed a correlation between concentration of antibiotics in the sample and intensity of fluorescent signal.

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
    Illegal NheI site found at 2888
    Illegal NotI site found at 579
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 588
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 2493
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 323
    Illegal BsaI.rc site found at 1919