Difference between revisions of "Part:BBa K2486010"
 
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This module works as a reporter for lactate an tetracycline presence. The circuit's key part is the PlldR_tetR promoter ([https://parts.igem.org/wiki/index.php?title=Part:BBa_K2486011 BBa_K2486011]), a part inspired by ETH_Zurich 2015 team [http://2015.igem.org/Team:ETH_Zurich/Part_Collection parts collection] of hybrid promoters for LacI and LldR. We adopted tetR operator in our promoter instead of lacI operator because of it's tighter regulation, reducing noise and basal signal of the system. The other components are:
 
This module works as a reporter for lactate an tetracycline presence. The circuit's key part is the PlldR_tetR promoter ([https://parts.igem.org/wiki/index.php?title=Part:BBa_K2486011 BBa_K2486011]), a part inspired by ETH_Zurich 2015 team [http://2015.igem.org/Team:ETH_Zurich/Part_Collection parts collection] of hybrid promoters for LacI and LldR. We adopted tetR operator in our promoter instead of lacI operator because of it's tighter regulation, reducing noise and basal signal of the system. The other components are:
 
# [https://parts.igem.org/Part:BBa_B0034 BBa_B0034]: a strong RBS from [the https://parts.igem.org/Ribosome_Binding_Sites/Prokaryotic/Constitutive/Community_Collection Community Collection]
 
# [https://parts.igem.org/Part:BBa_B0034 BBa_B0034]: a strong RBS from [the https://parts.igem.org/Ribosome_Binding_Sites/Prokaryotic/Constitutive/Community_Collection Community Collection]
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#  
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# [https://parts.igem.org/wiki/index.php?title=Part:BBa_K2486024 BBa_K2486024]: a 6 base pair spacer for optimal expression.
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# [https://parts.igem.org/wiki/index.php?title=Part:BBa_K082003 BBa_K082003]: a GFP with LVA tag
  
 
===Characterization===
 
===Characterization===
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https://static.igem.org/mediawiki/2017/0/0d/T--USP-Brazil--Detecttion_Iron_Lactate_concept_%285%29.PNG
 
https://static.igem.org/mediawiki/2017/0/0d/T--USP-Brazil--Detecttion_Iron_Lactate_concept_%285%29.PNG
  
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The data of the validation of the parts in the presence of TetR and LldR are described in parts [https://parts.igem.org/wiki/index.php?title=Part:BBa_K2486019 BBa_K2486019] (LldR-based lactate biosensor) and [https://parts.igem.org/wiki/index.php?title=Part:BBa_K2486020 BBa_K2486020] (TetR-based tetracycline biosensor) and here.
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The data of the validation of this part in the presence of LldR and TetR is described in parts [https://parts.igem.org/wiki/index.php?title=Part:BBa_K2486019 BBa_K2486019] (LldR-based lactate biosensor) and [https://parts.igem.org/wiki/index.php?title=Part:BBa_K2486020 BBa_K2486020] (TetR-based tetracycline biosensor) and [https://parts.igem.org/Part:BBa_K2486010 here].
  
 
===PlldR_tetR promoter activity in the presence of LldR regulator===
 
===PlldR_tetR promoter activity in the presence of LldR regulator===
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===PlldR_tetR promoter activity in the presence of TetR regulator===
 
===PlldR_tetR promoter activity in the presence of TetR regulator===
  
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https://static.igem.org/mediawiki/2017/thumb/8/86/T--USP-Brazil--Detecttion_Iron_Lactate_TetR%281%29.png/800px-T--USP-Brazil--Detecttion_Iron_Lactate_TetR%281%29.png
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https://static.igem.org/mediawiki/2017/0/04/T--USP-Brazil--Detecttion_Iron_Lactate_TetR%282%29.png
  
 
===Conclusion===
 
===Conclusion===
  
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The presence of LlldR and lactate affects the circuit as expected, repressing it in the absence of lactate. The presence of TetR and tetracycline could not be adequately mesured probably due to the Tetracycline used, once Anhydrotetracycline was not available.
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The presence of LlldR and lactate affects the circuit as expected, repressing it in the absence of lactate. The presence of TetR and tetracycline worked as expected too, sugesting that the part could work with both regulators.
  
 
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Latest revision as of 03:40, 2 November 2017


PlldR_tetR reporter circuit (GFP)

This module works as a reporter for lactate an tetracycline presence. The circuit's key part is the PlldR_tetR promoter (BBa_K2486011), a part inspired by ETH_Zurich 2015 team [http://2015.igem.org/Team:ETH_Zurich/Part_Collection parts collection] of hybrid promoters for LacI and LldR. We adopted tetR operator in our promoter instead of lacI operator because of it's tighter regulation, reducing noise and basal signal of the system. The other components are:

  1. BBa_B0034: a strong RBS from [the https://parts.igem.org/Ribosome_Binding_Sites/Prokaryotic/Constitutive/Community_Collection Community Collection]
  2. BBa_K2486024: a 6 base pair spacer for optimal expression.
  3. BBa_K082003: a GFP with LVA tag

Characterization

T--USP-Brazil--Detecttion_Iron_Lactate_concept_%285%29.PNG

The data of the validation of this part in the presence of LldR and TetR is described in parts BBa_K2486019 (LldR-based lactate biosensor) and BBa_K2486020 (TetR-based tetracycline biosensor) and here.

PlldR_tetR promoter activity in the presence of LldR regulator

800px-T--USP-Brazil--Detecttion_Iron_Lactate_lldR%281%29.png

PlldR_tetR promoter activity in the presence of TetR regulator

T--USP-Brazil--Detecttion_Iron_Lactate_TetR%282%29.png

Conclusion

The presence of LlldR and lactate affects the circuit as expected, repressing it in the absence of lactate. The presence of TetR and tetracycline worked as expected too, sugesting that the part could work with both regulators.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 100
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 810