Difference between revisions of "Part:BBa K3793004"
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<h2>MilkClear iGEM UCopenhagen 2024</h2> | <h2>MilkClear iGEM UCopenhagen 2024</h2> | ||
| − | https://2024.igem.wiki/ucopenhagen/ | + | Author: Kate Escobar, 26-09-2024, MilkClear's Wiki: https://2024.igem.wiki/ucopenhagen/ |
This part was used in our biosensor system to detect polycyclic aromatic hydrocarbons (PAHs), dioxin-like polychlorinated biphenyls (PCBs) and dioxin in breast milk. More information about our the composite part and device where AhR is included are linked here: [Paste the links]. | This part was used in our biosensor system to detect polycyclic aromatic hydrocarbons (PAHs), dioxin-like polychlorinated biphenyls (PCBs) and dioxin in breast milk. More information about our the composite part and device where AhR is included are linked here: [Paste the links]. | ||
Revision as of 14:49, 26 September 2024
CDS - Human Aryl hydrocarbon Receptor (Truncated)
This sequence encodes for the truncated version of the human Aryl hydrocarbon receptor (AhR). AhR is a transcription factor that is activated by ligands such as the 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD). Tryptophan metbolites such as kynurenic acid form an important class of ligands to the AhR. Upon Ligand binding, AhR translocates to the nucleus where it forms a heterodimer with the AhR Nuclear Translocator (ARNT). The heterodimer binds to the DNA sequence 5'-TGCGTG-3' within the dioxin response element (DRE) of target gene promoters and activates their transcription.
The part contains the coding sequence for a truncated version of AhR containing the the basic helix-loop-helix, PAS-A and PAS-B domains. The CDS has been codon optimized to remove any type IIS restriction enzyme sites. An N-Terminal 6x HIS tag is added to the CDS.
MilkClear iGEM UCopenhagen 2024
Author: Kate Escobar, 26-09-2024, MilkClear's Wiki: https://2024.igem.wiki/ucopenhagen/
This part was used in our biosensor system to detect polycyclic aromatic hydrocarbons (PAHs), dioxin-like polychlorinated biphenyls (PCBs) and dioxin in breast milk. More information about our the composite part and device where AhR is included are linked here: [Paste the links].
In its inactive state, AhR is located in the cytosol, where it is protected by HSP90 from degradation and nuclear translocation (1). Upon binding these ligands, AhR1 undergoes a conformational change that allows it to translocate to the nucleus (2), where it dimerizes with the Aryl hydrocarbon receptor nuclear translocator (ARNT) (1). This complex binds to specific DNA sequences known as xenobiotic response elements (XREs) and therefore allows for transcription of NanoLuc.
Our findings from Drylab
[Paste it here]
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 364
- 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 364
Illegal NheI site found at 169 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 364
Illegal BglII site found at 1179 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 364
- 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 364
- 1000COMPATIBLE WITH RFC[1000]
References
1. Carambia, A., Schuran, F.A. The aryl hydrocarbon receptor in liver inflammation. Semin Immunopathol 43, 563–575 (2021). https://doi.org/10.1007/s00281-021-00867-8
2. Mandal A, Biswas N, Alam MN. Implications of xenobiotic-response element(s) and aryl hydrocarbon receptor in health and diseases. Hum Cell. 2023 Sep;36(5):1638-1655. doi: 10.1007/s13577-023-00931-5. Epub 2023 Jun 17. PMID: 37329424.