Part:BBa_K4442000
Indole Inducible System
The indole-inducible sensing system PpTrpI/PPP_RS00425 originates from Pseudomonas Putida KT2440 which was identified and characterized by Matulis et al. (2022). The system below includes the nucleotide sequence containing the trpI gene, encoding the LysR family transcriptional regulator (locus tag PP_RS00430), and intergenic region PP_RS00430/PP_RS00425, assembled in construct pPM0081. The system exhibits a strong affinity for indole, which activates the system, and is strictly specific. A fluorescent reporter or any other output protein can be added after the promoter region at the end of the sequence, just be sure to add an RBS directly after the promoter.
More information can be found here:
Matulis P, Kutraite I, Augustiniene E, Valanciene E, Jonuskiene I, Malys N. Development and Characterization of Indole-Responsive Whole-Cell Biosensor Based on the Inducible Gene Expression System from Pseudomonas putida KT2440. International Journal of Molecular Sciences. 2022; 23(9):4649. https://doi.org/10.3390/ijms23094649
In literature, an increase in indole concentration in fecal samples was found to be correlated with depressive symptoms. In our project, we aimed to create a microbial sensor of the gut to aid in the diagnosis of Major Depressive Disorder (MDD), and by measuring fluorescence intensity, this system will allow us to detect the levels of indole.
Although we were unable to test our novel system in the wet lab, the article referenced above by Matulis et al. (2022) presented results on the fluorescence detected in response to different concentrations of indole.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 154
Illegal NgoMIV site found at 212 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 504
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