Difference between revisions of "Part:BBa K3777025"
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<br>Compared to our another part ctcS-tetM-T7(ctcO)-3WJdB(BBa_K3777018),it adds a binding site which can be regonized by dcas9 proteins and sgRNA.(Fig 1)  
 
<br>Compared to our another part ctcS-tetM-T7(ctcO)-3WJdB(BBa_K3777018),it adds a binding site which can be regonized by dcas9 proteins and sgRNA.(Fig 1)  
 
<br>When CTR was absent, ctcs 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. But BS2 will be regonized and cut by dca9 proteins and sgRNA leading to lower fluorescence expression.  
 
<br>When CTR was absent, ctcs 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. But BS2 will be regonized and cut by dca9 proteins and sgRNA leading to lower fluorescence expression.  
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<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/d/d9/KB2-CtcS-T7%28ctcO%29-3WJdB-PctcS-sgRNA.PNG/798px-KB2-CtcS-T7%28ctcO%29-3WJdB-PctcS-sgRNA.PNG
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Chlortetracycline
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<br>                          Fig.1  Schematic overview of the genetic circuit.
 
<br>                          Fig.1  Schematic overview of the genetic circuit.
  

Revision as of 18:49, 21 October 2021


KB2-CtcS-T7(ctcO)-3WJdB-PctcS-sgRNA

Basic biosensor device for Chlortetracycline detection.

Usage and Biology
Compared to our another part ctcS-tetM-T7(ctcO)-3WJdB(BBa_K3777018),it adds a binding site which can be regonized by dcas9 proteins and sgRNA.(Fig 1)
When CTR was absent, ctcs 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. But BS2 will be regonized and cut by dca9 proteins and sgRNA leading to lower fluorescence expression. 798px-KB2-CtcS-T7%28ctcO%29-3WJdB-PctcS-sgRNA.PNG
Fig.1 Schematic overview of the genetic circuit.


Reference：Alam Khalid K,Tawiah Kwaku D,Lichte Matthew F,Porciani David,Burke Donald H. A Fluorescent Split Aptamer for Visualizing RNA-RNA Assembly In Vivo.[J]. ACS synthetic biology,2017,6(9)
Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 260
    Illegal NheI site found at 283
    Illegal NheI site found at 1164
    Illegal NotI site found at 832
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 121
    Illegal XhoI site found at 841
    Illegal XhoI site found at 1305
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 576