Difference between revisions of "Part:BBa K5477026"
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<partinfo>BBa_K5477026 short</partinfo> | <partinfo>BBa_K5477026 short</partinfo> | ||
− | The pRET2-NCOA composite part consists of the NCOA (Nuclear Receptor Coactivator) [https://parts.igem.org/Part:BBa_K5477012 | + | The pRET2-NCOA composite part consists of the NCOA (Nuclear Receptor Coactivator) [https://parts.igem.org/Part:BBa_K5477012 BBa_K5477012] gene under the control of the pRET2 promoter [https://parts.igem.org/Part:BBa_K5477000 BBa_K5477000], creating a module that enhances transcriptional activation by nuclear receptors (3) (4). NCOA, also known as SRC (Steroid Receptor Coactivator), is a coactivator protein that interacts with activated nuclear receptors to promote transcription of target genes. |
− | This module supports the AhR-mediated response to toxic ligands like PAHs, dioxins and PCBs by binding to the XRE | + | This module supports the AhR-mediated response to toxic ligands like PAHs, dioxins and PCBs by binding to the XRE in our reporter module facilitating the transcription of NanoLuc [https://parts.igem.org/Part:BBa_K5477030 BBa_K5477030] (1) (2). The composite part was cloned using the method of USER-cloning into YCp-W. YCp-W is a centromeric plasmid used in yeast that carries a TRP1 marker, allowing for selection in tryptophan auxotrophic yeast strains. Like other CEN plasmids, YCp-W contains a CEN sequence, ensuring that it replicates and segregates similarly to yeast chromosomes. This low-copy plasmid (typically maintained at one to two copies per cell) provides stable maintenance within yeast cells. This composite part was used in our device: [https://parts.igem.org/Part:BBa_K5477041 BBa_K5477041] and [https://parts.igem.org/Part:BBa_K5477042 BBa_K5477042] |
Latest revision as of 00:57, 2 October 2024
pRET2-NCOA - receptor module
The pRET2-NCOA composite part consists of the NCOA (Nuclear Receptor Coactivator) BBa_K5477012 gene under the control of the pRET2 promoter BBa_K5477000, creating a module that enhances transcriptional activation by nuclear receptors (3) (4). NCOA, also known as SRC (Steroid Receptor Coactivator), is a coactivator protein that interacts with activated nuclear receptors to promote transcription of target genes.
This module supports the AhR-mediated response to toxic ligands like PAHs, dioxins and PCBs by binding to the XRE in our reporter module facilitating the transcription of NanoLuc BBa_K5477030 (1) (2). The composite part was cloned using the method of USER-cloning into YCp-W. YCp-W is a centromeric plasmid used in yeast that carries a TRP1 marker, allowing for selection in tryptophan auxotrophic yeast strains. Like other CEN plasmids, YCp-W contains a CEN sequence, ensuring that it replicates and segregates similarly to yeast chromosomes. This low-copy plasmid (typically maintained at one to two copies per cell) provides stable maintenance within yeast cells. This composite part was used in our device: BBa_K5477041 and BBa_K5477042
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 1785
Illegal EcoRI site found at 2407
Illegal SpeI site found at 1234
Illegal SpeI site found at 1535
Illegal SpeI site found at 2602
Illegal PstI site found at 1294 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 1785
Illegal EcoRI site found at 2407
Illegal NheI site found at 182
Illegal NheI site found at 3547
Illegal SpeI site found at 1234
Illegal SpeI site found at 1535
Illegal SpeI site found at 2602
Illegal PstI site found at 1294 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 1785
Illegal EcoRI site found at 2407
Illegal BamHI site found at 3049 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 1785
Illegal EcoRI site found at 2407
Illegal SpeI site found at 1234
Illegal SpeI site found at 1535
Illegal SpeI site found at 2602
Illegal PstI site found at 1294 - 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 1785
Illegal EcoRI site found at 2407
Illegal SpeI site found at 1234
Illegal SpeI site found at 1535
Illegal SpeI site found at 2602
Illegal PstI site found at 1294
Illegal AgeI site found at 37 - 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. Goedtke L, Sprenger H, Hofmann U, Schmidt FF, Hammer HS, Zanger UM, Poetz O, Seidel A, Braeuning A, Hessel-Pras S. Polycyclic Aromatic Hydrocarbons Activate the Aryl Hydrocarbon Receptor and the Constitutive Androstane Receptor to Regulate Xenobiotic Metabolism in Human Liver Cells. Int J Mol Sci. 2020 Dec 31;22(1):372. doi: 10.3390/ijms22010372. PMID: 33396476; PMCID: PMC7796163.
3. Onate SA, Boonyaratanakornkit V, Spencer TE, et al. The steroid receptor coactivator-1 contains multiple receptor interacting and activation domains that cooperatively enhance the activation function 1 (AF1) and AF2 domains of steroid receptors. J Biol Chem. 1998;273(20):12101-12108. doi:10.1074/jbc.273.20.12101
4. Oñate SA, Tsai SY, Tsai MJ, O'Malley BW. Sequence and characterization of a coactivator for the steroid hormone receptor superfamily. Science. 1995;270(5240):1354-1357. doi:10.1126/science.270.5240.1354