Difference between revisions of "Part:BBa M45699"

Latest revision as of 16:17, 23 April 2015

Vitamin D receptor with red chromoprotein reporter

This system consists of two parts: Promoter with Vitamin D receptor hinge region and chromoprotein eforRed. the Vitamin D bonds to the receptor and activate the express of the chromoprotein eforRed. The red color can be observed when the presence of Vitamin D in the culture.

Estrogen receptor DBD - Vitamin D receptor hinge region - Estrogen receptor LBD Artificial eukaryotic TF made of Estrogen receptor's DBD (DNA Binding Domain), a vitamin D3 receptor hinge domain and H. This chromoprotein eforRed naturally exhibits red/pink color when expressed. The color is slightly weaker than RFP. On agar plates and in liquid culture, the color is readily visible to naked eye in less than 24 hours of incubation. The DNA was codon-optimized for expression in E.coli and synthesized by the Korean company Bioneer Corporation.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

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 __NOTOC__
 <partinfo>BBa_M45699 short</partinfo>
+This system consists of two parts: Promoter with Vitamin D receptor hinge region and chromoprotein eforRed. the Vitamin D bonds to the receptor and activate the express of the chromoprotein eforRed. The red color can be observed when the presence of Vitamin D in the culture.
-Estrogen receptor DBD - Vitamin D receptor hinge region - Estrogen receptor LBD
+Estrogen receptor DBD - Vitamin D receptor hinge region - Estrogen receptor LBD Artificial eukaryotic TF made of Estrogen receptor's DBD (DNA Binding Domain), a vitamin D3 receptor hinge domain and H.
-Artificial eukaryotic TF made of Estrogen receptor's DBD (DNA Binding Domain), a vitamin D3 receptor hinge domain and H. sapiens nuclear hormone receptor LBD (Ligand Binding Domain)
-Human Estrogen Receptor
-ER-&#945; is a 17&#946;-estradiol-activated steroid receptor member of the nuclear receptor superfamily of transcription factors. It has a variety of central physiological roles, including those involved in maintenance of the reproductive, cardiovascular, musculoskeletal and central nervous systems. ER-&#945; is expressed at low to moderate levels in major physiological systems (central nervous system (CNS), endocrine, metabolic, gastrointestinal, immune, reproductive, cardiovascular, respiratory and structural), with peaks of expression in the pituitary, ovary, uterus and vas deferens. ER-&#945; dysfunction is associated with cancer, cardiovascular system defects, hematological system defects, immune and inflammation diseases, metabolic defects, reproductive defects.
-This composite artificial transcription factor will activate any reporter or any gene in general that has a UAS (Upper Activating Sequence) 3' of its promoter. The usual binding sites of reporters, contain multiple UAS elements. In order to have a POPS output, the LBD has to recruit activators in the cell. This can be initiated by ligand binding or by recruiting a protein that has a fused strong activator like the VP activator.
-With this system NHR (Nuclear Hormone Receptor) ligands or NHR interacting partners can be screened.
-The NHR: cofactor-VP interaction should be also broken by a potential ligand binding, this is why this setup is also suitable for ligand identification. The benefit of the cofactor-VP interaction test is that the dynamic range of the assay is much higher than the dynamic range of the normal Gal4-NHR ligand activation assay.
-More info about this project on the wiki pages of Team Debrecen-Hungary 2010 [1]
 This chromoprotein eforRed naturally exhibits red/pink color when expressed. The color is slightly weaker than RFP. On agar plates and in liquid culture, the color is readily visible to naked eye in less than 24 hours of incubation. The DNA was codon-optimized for expression in E.coli and synthesized by the Korean company Bioneer Corporation.