Difference between revisions of "Part:BBa K2486010"

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:

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

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

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

@@ Line 24: / Line 24: @@
 ===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 to, sugesting that the part could work with both regulators.
+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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