Difference between revisions of "Part:BBa K5477023"
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− | The pSTE12-AhR receptor module is a composite | + | The pSTE12-AhR receptor module is designed as a composite with AhR [https://parts.igem.org/Part:BBa_K3793004 BBa_K3793004]is expressed under the control of the pSTE12 promoter [https://parts.igem.org/Part:BBa_K5477001 BBa_K5477001]. AhR is a ligand-activated transcription factor that binds to a variety of environmental contaminants, including polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs), and dioxins (1) (2). Upon binding these ligands, AhR translocates from the cytoplasm to the nucleus, where it forms a heterodimer with the Aryl hydrocarbon receptor nuclear translocator (ARNT). This complex then binds to xenobiotic response elements (XREs), initiating the transcription of detoxification genes, such as those encoding cytochrome P450 enzymes. |
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+ | In the pSTE12-AhR receptor module, the pSTE12 promoter drives the expression of AhR. Once AhR is expressed and activated by ligands like PAHs or PCBs, it binds to the XRE sequence in a reporter module, triggering the expression of NanoLuc. The composite part was cloned using the method of USER-cloning into YCp-L. The YCp-L plasmid is a centromeric plasmid that carries a LEU2 marker for leucine selection in yeast. Centromeric plasmids like YCp-L are low-copy vectors, typically maintained at one to two copies per cell due to the presence of a CEN sequence. This composite part was used in this device: [https://parts.igem.org/Part:BBa_K5477041 BBa_K5477041] | ||
Latest revision as of 00:40, 2 October 2024
pSTE12-AhR - receptor module
The pSTE12-AhR receptor module is designed as a composite with AhR BBa_K3793004is expressed under the control of the pSTE12 promoter BBa_K5477001. AhR is a ligand-activated transcription factor that binds to a variety of environmental contaminants, including polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs), and dioxins (1) (2). Upon binding these ligands, AhR translocates from the cytoplasm to the nucleus, where it forms a heterodimer with the Aryl hydrocarbon receptor nuclear translocator (ARNT). This complex then binds to xenobiotic response elements (XREs), initiating the transcription of detoxification genes, such as those encoding cytochrome P450 enzymes.
In the pSTE12-AhR receptor module, the pSTE12 promoter drives the expression of AhR. Once AhR is expressed and activated by ligands like PAHs or PCBs, it binds to the XRE sequence in a reporter module, triggering the expression of NanoLuc. The composite part was cloned using the method of USER-cloning into YCp-L. The YCp-L plasmid is a centromeric plasmid that carries a LEU2 marker for leucine selection in yeast. Centromeric plasmids like YCp-L are low-copy vectors, typically maintained at one to two copies per cell due to the presence of a CEN sequence. This composite part was used in this device: BBa_K5477041
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 927
Illegal XbaI site found at 55 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 927
Illegal NheI site found at 732 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 927
Illegal BglII site found at 1742 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 927
Illegal XbaI site found at 55 - 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 927
Illegal XbaI site found at 55 - 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.