Difference between revisions of "Part:BBa K3594003"

 
 
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This composite part is used as the reporter of the quorum sensing device. It consists a Plux promotor and BBa_K3594009, a sfGFP gene. When the concentration of the signal molecule-LuxR-AHL reaches threshold, the bacteria start to commit suicide and at the same time, emit large amount of fluorescence.
 
This composite part is used as the reporter of the quorum sensing device. It consists a Plux promotor and BBa_K3594009, a sfGFP gene. When the concentration of the signal molecule-LuxR-AHL reaches threshold, the bacteria start to commit suicide and at the same time, emit large amount of fluorescence.
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[[File:T--SHSBNU China--synchronized lysis circuit in M.Omar Din et al. Nature, 2016.jpg|500px|thumb|center|synchronized lysis circuit in M.Omar Din et al. Nature, 2016]]
  
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In our project, we made the following changes to the above experiment:
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[[File:T--SHSBNU China--the design of our bacterial synchronized lysis circuit.png|500px|thumb|center|the design of our bacterial synchronized lysis circuit]]
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pSHS6_pSB1C3_pTac_LuxR_plux_sfGFP: constructed by Gibson assembly
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T4 Ligation: pTac_LuxR_plux_sfGFP（~3.6Kb）and pSB1C3 backbone（~2Kb）
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This plasmid is used to test if LuxR and plux promoter work well.
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[[File:T--SHSBNU China--the electrophoretogram of colony PCR of BBa K3594003.png|500px|thumb|center|the electrophoretogram of colony PCR of BBa K3594003]]
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Verification of LuxR and Plux
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[[File:T--SHSBNU China--sequence result of BBa K3594003.png|500px|thumb|center|Sequencing results of plux-GFP]]
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[[File:T--SHSBNU China--BBa K3594003.png|500px|thumb|center|The result of BBa_K3594003]]
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If the fluorescence value in the tube increases with time, and the fluorescence value in the tube with IPTG and atc is higher than no inducer or only one inducer, it proves that the bacteria emit fluorescence after reaching the threshold, therefore the experiment is successful.
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here
 
===Usage and Biology===
 
===Usage and Biology===

Latest revision as of 19:15, 23 October 2020


Quorum Sensing Fluorescence Reporting Device

This composite part is used as the reporter of the quorum sensing device. It consists a Plux promotor and BBa_K3594009, a sfGFP gene. When the concentration of the signal molecule-LuxR-AHL reaches threshold, the bacteria start to commit suicide and at the same time, emit large amount of fluorescence.

synchronized lysis circuit in M.Omar Din et al. Nature, 2016

In our project, we made the following changes to the above experiment:

the design of our bacterial synchronized lysis circuit

pSHS6_pSB1C3_pTac_LuxR_plux_sfGFP: constructed by Gibson assembly T4 Ligation: pTac_LuxR_plux_sfGFP（~3.6Kb）and pSB1C3 backbone（~2Kb） This plasmid is used to test if LuxR and plux promoter work well.

the electrophoretogram of colony PCR of BBa K3594003

Verification of LuxR and Plux

Sequencing results of plux-GFP
The result of BBa_K3594003

If the fluorescence value in the tube increases with time, and the fluorescence value in the tube with IPTG and atc is higher than no inducer or only one inducer, it proves that the bacteria emit fluorescence after reaching the threshold, therefore the experiment is successful. Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 1208
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]