Difference between revisions of "Part:BBa K5477029"

Latest revision as of 20:15, 26 September 2024

pRET2-LexA-mERα(LBD) - receptor module

The pRET2-LexA-mERα(LBD) composite part is an receptor module that integrates the pRET2 promoter BBa_K5477000with a mutant version of the Estrogen Receptor Alpha ligand-binding domain (LBD), fused to the LexA DNA-binding domain (DBD)BBa_K5477014. This allows for the regulation of gene expression in response to bisphenol A (BPA) or other estrogen-like molecules, but with altered specificity and dynamics due to the mutation in the ERα LBD (3). In this system, the LexA- mERα(LBD) targets specific LexA operator sequences (Lex6Op) in our reporter module. The mutant ERα LBD (mERα) in this construct is designed to have altered ligand-binding properties compared to the wild-type ERα, allowing it to respond differently to ligands like BPA, which is known to act as an endocrine disruptor (1) (2) (3) (4) .

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:

Sequence and Features

Assembly Compatibility:

10
INCOMPATIBLE WITH RFC[10]
Illegal XbaI site found at 1606
Illegal PstI site found at 1790
Illegal PstI site found at 1961
12
INCOMPATIBLE WITH RFC[12]
Illegal NheI site found at 182
Illegal PstI site found at 1790
Illegal PstI site found at 1961
21
INCOMPATIBLE WITH RFC[21]
Illegal BglII site found at 1740
23
INCOMPATIBLE WITH RFC[23]
Illegal XbaI site found at 1606
Illegal PstI site found at 1790
Illegal PstI site found at 1961
25
INCOMPATIBLE 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
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI site found at 1640

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 α†‡. 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

@@ Line 3: / Line 3: @@
 <partinfo>BBa_K5477029 short</partinfo>
-The pRET2-LexA-mER&#945;(LBD) composite part is an receptor module that integrates the pRET2 promoter with a mutant version of the Estrogen Receptor Alpha ligand-binding domain (LBD), fused to the LexA DNA-binding domain (DBD). This fusion protein allows for the regulation of gene expression in response to bisphenol A (BPA) or other estrogen-like molecules, but with altered specificity and dynamics due to the mutation in the ER&#945; LBD. In this system, the LexA- mER&#945;(LBD) targets specific LexA operator sequences (Lex6Op) in our reporter module. The mutant ER&#945; LBD (mER&#945;) in this construct is designed to have altered ligand-binding properties compared to the wild-type ER&#945;, allowing it to respond differently to ligands like BPA, which is known to act as an endocrine disruptor.
+The pRET2-LexA-mER&#945;(LBD) composite part is an receptor module that integrates the pRET2 promoter [https://parts.igem.org/Part:BBa_K5477000| BBa_K5477000]with a mutant version of the Estrogen Receptor Alpha ligand-binding domain (LBD), fused to the LexA DNA-binding domain (DBD)[https://parts.igem.org/Part:BBa_K5477014| BBa_K5477014]. This allows for the regulation of gene expression in response to bisphenol A (BPA) or other estrogen-like molecules, but with altered specificity and dynamics due to the mutation in the ER&#945; LBD (3). In this system, the LexA- mER&#945;(LBD) targets specific LexA operator sequences (Lex6Op) in our reporter module. The mutant ER&#945; LBD (mER&#945;) in this construct is designed to have altered ligand-binding properties compared to the wild-type ER&#945;, allowing it to respond differently to ligands like BPA, which is known to act as an endocrine disruptor (1) (2) (3) (4) .
+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:
@@ Line 18: / Line 20: @@
 <partinfo>BBa_K5477029 parameters</partinfo>
 <!-- -->
+===References===
+. 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.
+. 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.
+. Rajasärkkä, J., Hakkila, K. and Virta, M. (2011), Developing a compound-specific receptor for bisphenol a by directed evolution of human estrogen receptor α†‡. 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
+. 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