Difference between revisions of "Part:BBa K3777023"
 
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<partinfo>BBa_K3777023 short</partinfo>
 
<partinfo>BBa_K3777023 short</partinfo>
  
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Basic biosensor device for tetracycline detection.
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Basic biosensor device.
  
 
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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 curcuit includes 3WJdB(BBa_K3777000) which is under the control of T7 promoter (BBa_K3777006).  The terminator we used was 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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https://static.igem.org/mediawiki/parts/9/94/T7-3WJdB%E2%91%A1.PNG
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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/3/3d/Tetr-tetO-3WJdB.PNG/799px-Tetr-tetO-3WJdB.PNG  
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<br>                          Fig.1 Schematic overview of the genetic circuit.
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<br><b><font size="3">Results</font></b>
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<br>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.
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Latest revision as of 17:34, 21 October 2021


T7-3WJdB

Basic biosensor device.

Usage and Biology
The curcuit includes 3WJdB(BBa_K3777000) which is under the control of T7 promoter (BBa_K3777006). The terminator we used was BBa_M36305.(Fig 1) T7-3WJdB%E2%91%A1.PNG


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 206
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]