Difference between revisions of "Part:BBa K5477029"
Kateesc1700 (Talk | contribs) |
Kateesc1700 (Talk | contribs) |
||
(2 intermediate revisions by the same user not shown) | |||
Line 3: | Line 3: | ||
<partinfo>BBa_K5477029 short</partinfo> | <partinfo>BBa_K5477029 short</partinfo> | ||
− | |||
− | 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 | + | The pRET2-LexA-mERα(LBD) composite part integrates the LexA DNA-binding domain (DBD) with the engineered ligand-binding domain (LBD) of Estrogen Receptor Alpha (mERα) [https://parts.igem.org/Part:BBa_K5477014 BBa_K5477014] from the study of Rajasärkkä et al. 2011, under the control of the pRET2 promoter [https://parts.igem.org/Part:BBa_K5477000| BBa_K5477000]. This allows for the regulation of gene expression in response to estrogen or estrogen-like molecules, such as bisphenol A (BPA) (1) (2) (3) (4). |
+ | |||
+ | |||
+ | Upon presence of BPA, the LexA-ERα(LBD) binds specifically to LexA operator sequences (Lex6Op) which is a part of the reporter module that initiates transcription 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 the following devices: [https://parts.igem.org/Part:BBa_K5477045 BBa_K5477045] and [https://parts.igem.org/Part:BBa_K5477046 BBa_K5477046]. | ||
Latest revision as of 01:16, 2 October 2024
pRET2-LexA-mERα(LBD) - receptor module
The pRET2-LexA-mERα(LBD) composite part integrates the LexA DNA-binding domain (DBD) with the engineered ligand-binding domain (LBD) of Estrogen Receptor Alpha (mERα) BBa_K5477014 from the study of Rajasärkkä et al. 2011, under the control of the pRET2 promoter BBa_K5477000. This allows for the regulation of gene expression in response to estrogen or estrogen-like molecules, such as bisphenol A (BPA) (1) (2) (3) (4).
Upon presence of BPA, the LexA-ERα(LBD) binds specifically to LexA operator sequences (Lex6Op) which is a part of the reporter module that initiates transcription 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 the following devices: BBa_K5477045 and BBa_K5477046.
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 NheI site found at 182
Illegal PstI site found at 1790
Illegal PstI site found at 1961 - 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 1740
- 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
Illegal AgeI site found at 37 - 1000COMPATIBLE WITH RFC[1000]
References
1. Çiftçi S, Yalçın SS, Samur G. Bisphenol A Exposure in Exclusively Breastfed Infants and Lactating Women: An Observational Cross-sectional Study. J Clin Res Pediatr Endocrinol. 2021 Nov 25;13(4):375-383. doi: 10.4274/jcrpe.galenos.2020.2021.0305. Epub 2021 Mar 22. PMID: 33749218; PMCID: PMC8638632.
2. 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.
3. Rajasärkkä, J., Hakkila, K. and Virta, M. (2011), Developing a compound-specific receptor for bisphenol a by directed evolution of human estrogen receptor ᆇ. Biotechnol. Bioeng., 108: 2526-2534. https://doi.org/10.1002/bit.23214 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
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