Part:BBa_K1355002
Mercury ions detector device
We designed a biobrick device to express Green Fluorescent Protein in mercury’s occurrence. The Mercury ions’ detector device biobrick (BBa_K1355002) is composed by mer bidirectional promoter (BBa_K1355001) attached to the GFP translational unit (BBa_E0840) available on iGEM database since 2004 . It has dual function: A) In reverse: MerR protein regulator transcription; and B) in forward: transcription of MerP - MerT - GFP proteins, as represented below:
In absence of mercury, MerR forms a MerR-promoter-operator complex, preventing RNA polymerase to recognize the promoter, consequently, messengers RNA for MerPT and GFP will not be transcript. In presence of Hg2+, MerR protein binds to this element and dissociates from the promoter-operator complex, allowing MerPT and GFP expression, as represented below:
When MerT, MerP and the Green Fluorescent Protein (GFP) protein are expressed, the biodetection will start! MerP and MerT proteins are responsible for the transport of mercury from the periplasm to cytoplasm and GFP will act as a reporter alerting the presence of mercury in the middle and higher is mercury’s concentration, higher will be GFP expression.
Figure 1: Mercury Bacter Hg biodetector (DH5-alpha transformed with BBa_K1355002)
References
Farias, L. A., Fávaro, D. I., Pessoa, A., Aguiar, J. P., & Yuyama, L. K. (2012). Mercury and methylmercury concentration assessment in children's hair from Manaus, Amazonas state, Brazil. Acta Amazonica, 42(2), 279-286.
Fillion, M., Philibert, A., Mertens, F., Lemire, M., Passos, C. J. S., Frenette, B., ... & Mergler, D. (2011). Neurotoxic sequelae of mercury exposure: an intervention and follow-up study in the Brazilian Amazon. Ecohealth, 8(2), 210-222.
Grotto, D., Valentini, J., Fillion, M., Passos, C. J. S., Garcia, S. C., Mergler, D., & Barbosa Jr, F. (2010). Mercury exposure and oxidative stress in communities of the Brazilian Amazon. Science of the Total Environment, 408(4), 806-811.
Hakkila, K., Maksimow, M., Karp, M., & Virta, M. (2002). Reporter Genes lucFF, luxCDABE, gfp, and dsred Have Different Characteristics in Whole-Cell Bacterial Sensors. Analytical biochemistry, 301(2), 235-242.
Kuncova, G., Pazlarova, J., Hlavata, A., Ripp, S., & Sayler, G. S. (2011). Bioluminescent bioreporters Pseudomonas putida TVA8 as a detector of water pollution. Operational conditions and selectivity of free cells sensor. Ecological Indicators, 11(3), 882-887.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 988
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 586
Illegal NgoMIV site found at 1160 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 1862
Illegal SapI site found at 579
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