Difference between revisions of "Part:BBa K5477028"
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The pPOP6-LexA-ERα(LBD) composite part is a fusion module that combines the pPOP6 promoter [https://parts.igem.org/Part:BBa_K5477007 | BBa_K5477007] with the LexA-ERα(LBD) construct [https://parts.igem.org/Part:BBa_K5477013 | BBa_K5477013]. In this system, the pPOP6 promoter drives the expression of the LexA-ERα(LBD) fusion protein, which consists of the LexA DNA-binding domain (DBD) fused to the ligand-binding domain (LBD) of Estrogen Receptor Alpha (ERα). This design allows for precise ligand-dependent regulation of our reporter module based on the presence of estrogen or estrogen-like compounds like BPA (1) (2) (3) (4). Upon binding to the LexA-ERα(LBD), this acts as a transcription factor and binds to LexA operator sequences (Lex6Op) present in our reporter module expressing NanoLuc. | The pPOP6-LexA-ERα(LBD) composite part is a fusion module that combines the pPOP6 promoter [https://parts.igem.org/Part:BBa_K5477007 | BBa_K5477007] with the LexA-ERα(LBD) construct [https://parts.igem.org/Part:BBa_K5477013 | BBa_K5477013]. In this system, the pPOP6 promoter drives the expression of the LexA-ERα(LBD) fusion protein, which consists of the LexA DNA-binding domain (DBD) fused to the ligand-binding domain (LBD) of Estrogen Receptor Alpha (ERα). This design allows for precise ligand-dependent regulation of our reporter module based on the presence of estrogen or estrogen-like compounds like BPA (1) (2) (3) (4). Upon binding to the LexA-ERα(LBD), this acts as a transcription factor and binds to LexA operator sequences (Lex6Op) present in our reporter module expressing 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 our device: | + | 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 our device [https://parts.igem.org/Part:BBa_K5477044 BBa_K5477044] |
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Revision as of 19:45, 29 September 2024
pPOP6-LexA-ERα(LBD) - receptor module
The pPOP6-LexA-ERα(LBD) composite part is a fusion module that combines the pPOP6 promoter | BBa_K5477007 with the LexA-ERα(LBD) construct | BBa_K5477013. In this system, the pPOP6 promoter drives the expression of the LexA-ERα(LBD) fusion protein, which consists of the LexA DNA-binding domain (DBD) fused to the ligand-binding domain (LBD) of Estrogen Receptor Alpha (ERα). This design allows for precise ligand-dependent regulation of our reporter module based on the presence of estrogen or estrogen-like compounds like BPA (1) (2) (3) (4). Upon binding to the LexA-ERα(LBD), this acts as a transcription factor and binds to LexA operator sequences (Lex6Op) present in our reporter module expressing 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 our device BBa_K5477044
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal XbaI site found at 1606
Illegal PstI site found at 1790
Illegal PstI site found at 1961 - 12INCOMPATIBLE WITH RFC[12]Illegal PstI site found at 1790
Illegal PstI site found at 1961 - 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 1740
Illegal BamHI site found at 73
Illegal BamHI site found at 390 - 23INCOMPATIBLE WITH RFC[23]Illegal XbaI site found at 1606
Illegal PstI site found at 1790
Illegal PstI site found at 1961 - 25INCOMPATIBLE WITH RFC[25]Illegal XbaI site found at 1606
Illegal PstI site found at 1790
Illegal PstI site found at 1961 - 1000COMPATIBLE WITH RFC[1000]
References
1. Hafezi SA, Abdel-Rahman WM. The Endocrine Disruptor Bisphenol A (BPA) Exerts a Wide Range of Effects in Carcinogenesis and Response to Therapy. Curr Mol Pharmacol. 2019;12(3):230-238. doi: 10.2174/1874467212666190306164507. PMID: 30848227; PMCID: PMC6864600.
2. Lo, Elena & Piparo, Elena & Siragusa, Lydia & Raymond, Frédéric & Passeri, Giovanna & Cruciani, Gabriele & Schilter, Benoit. (2019). Bisphenol A Binding Promiscuity: A Virtual Journey through the Universe of Proteins 1.
3. Park, Choa & Song, Heewon & Choi, Junyeong & Sim, Seunghye & Kojima, Hiroyuki & Park, Joonwoo & Iida, Mitsuru & Lee, Youngjoo. (2020). The mixture effects of bisphenol derivatives on estrogen receptor and androgen receptor. Environmental Pollution. 260. 114036. 10.1016/j.envpol.2020.114036.
4. Zhou, T., Liang, Z. & Marchisio, M.A. Engineering a two-gene system to operate as a highly sensitive biosensor or a sharp switch upon induction with β-estradiol. Sci Rep 12, 21791 (2022). https://doi.org/10.1038/s41598-022-26195-x