Difference between revisions of "Part:BBa K2486010"
Line 12: | Line 12: | ||
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 | ||
− | The data of the validation of this part in the presence of | + | 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=== |
Revision as of 02:54, 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:
- BBa_B0034: a strong RBS from [the https://parts.igem.org/Ribosome_Binding_Sites/Prokaryotic/Constitutive/Community_Collection Community Collection]
- BBa_K2486024: a 6 base pair spacer for optimal expression.
- BBa_K082003: a GFP with LVA tag
Characterization
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
PlldR_tetR promoter activity in the presence of TetR regulator
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 could not be adequately mesured probably due to the Tetracycline used, once Anhydrotetracycline was not available.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 100
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 810