Difference between revisions of "Part:BBa K5477028"

Revision as of 20:08, 26 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:

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 PstI site found at 1790
Illegal PstI site found at 1961
21
INCOMPATIBLE WITH RFC[21]
Illegal BglII site found at 1740
Illegal BamHI site found at 73
Illegal BamHI site found at 390
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
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI site found at 1640

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

@@ Line 3: / Line 3: @@
 <partinfo>BBa_K5477028 short</partinfo>
-The pPOP6-LexA-ER&#945;(LBD) composite part is a fusion module that combines the pPOP6 promoter with the LexA-ER&#945;(LBD) construct. In this system, the pPOP6 promoter drives the expression of the LexA-ER&#945;(LBD) fusion protein, which consists of the LexA DNA-binding domain (DBD) fused to the ligand-binding domain (LBD) of Estrogen Receptor Alpha (ER&#945;). This design allows for precise ligand-dependent regulation of our reporter module based on the presence of estrogen or estrogen-like compounds like BPA. Upon binding to the LexA-ER&#945;(LBD), this acts as a transcription factor and binds to LexA operator sequences (Lex6Op) present in our reporter module expressing NanoLuc.
+The pPOP6-LexA-ER&#945;(LBD) composite part is a fusion module that combines the pPOP6 promoter [https://parts.igem.org/Part:BBa_K5477007 | BBa_K5477007]
+ with the LexA-ER&#945;(LBD) construct [https://parts.igem.org/Part:BBa_K5477013 | BBa_K5477013]. In this system, the pPOP6 promoter drives the expression of the LexA-ER&#945;(LBD) fusion protein, which consists of the LexA DNA-binding domain (DBD) fused to the ligand-binding domain (LBD) of Estrogen Receptor Alpha (ER&#945;). 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-ER&#945;(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:
 <!-- Add more about the biology of this part here
@@ Line 17: / Line 20: @@
 <partinfo>BBa_K5477028 parameters</partinfo>
 <!-- -->
+===References===
+. 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.
+. 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.
+. 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.
+. 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