Team:Debrecen-Hungary/parts
From 2010.igem.org
(→Gal4-NHR23 Chimeric nuclear receptor) |
|||
(17 intermediate revisions not shown) | |||
Line 26: | Line 26: | ||
Our lipid detecting tool kit comprosis of 19 basic parts (11 ligand binding domains, 3 DNA binding domains and 5 other basic parts),9 composite chimeric receptors and 3 plasmid expression vectors and associated intermediates. Amongst our kit, parts of various speicies of the animalia kingdom may be found. The composite receptors are proof of concept that the basic parts acctually work. The bulk of the data (sequance, sub domains, 3d structure, ligand affinity and more..) can be found in the parts registry. | Our lipid detecting tool kit comprosis of 19 basic parts (11 ligand binding domains, 3 DNA binding domains and 5 other basic parts),9 composite chimeric receptors and 3 plasmid expression vectors and associated intermediates. Amongst our kit, parts of various speicies of the animalia kingdom may be found. The composite receptors are proof of concept that the basic parts acctually work. The bulk of the data (sequance, sub domains, 3d structure, ligand affinity and more..) can be found in the parts registry. | ||
+ | |||
+ | <html><br><a name="LBD"></a><br></html> | ||
==Ligand binding domains - Basic Parts == | ==Ligand binding domains - Basic Parts == | ||
Line 71: | Line 73: | ||
ER-α is a 17β-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-α 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-α dysfunction is associated with cancer, cardiovascular system defects, hematological system defects, immune and inflammation diseases, metabolic defects, reproductive defects. | ER-α is a 17β-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-α 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-α dysfunction is associated with cancer, cardiovascular system defects, hematological system defects, immune and inflammation diseases, metabolic defects, reproductive defects. | ||
[http://partsregistry.org/wiki/index.php?title=Part:BBa_K364309 Go to the official parts registry page for this part] | [http://partsregistry.org/wiki/index.php?title=Part:BBa_K364309 Go to the official parts registry page for this part] | ||
+ | |||
+ | <html><br><a name="DBD"></a><br></html> | ||
==DNA binding domains - Basic Parts== | ==DNA binding domains - Basic Parts== | ||
Line 83: | Line 87: | ||
Estrogen receptor refers to a group of receptors that are activated by the hormone 17β-estradiol[1] (estrogen) , which is a member of the nuclear hormone family of intracellular receptors. The main function of the estrogen receptor is as a DNA-binding transcription factor that regulates gene expression. | Estrogen receptor refers to a group of receptors that are activated by the hormone 17β-estradiol[1] (estrogen) , which is a member of the nuclear hormone family of intracellular receptors. The main function of the estrogen receptor is as a DNA-binding transcription factor that regulates gene expression. | ||
[http://partsregistry.org/wiki/index.php?title=Part:BBa_K364307 Go to the official parts registry page for this part] | [http://partsregistry.org/wiki/index.php?title=Part:BBa_K364307 Go to the official parts registry page for this part] | ||
+ | |||
+ | <html><br><a name="other"></a><br></html> | ||
==Other Basic Parts== | ==Other Basic Parts== | ||
Line 106: | Line 112: | ||
=== [http://partsregistry.org/wiki/index.php?title=Part:BBa_K364328 Gal4-DAF12 Chimeric nuclear receptor] === | === [http://partsregistry.org/wiki/index.php?title=Part:BBa_K364328 Gal4-DAF12 Chimeric nuclear receptor] === | ||
- | Artificial eukaryotic TF made of Gal4 DBD (DNA Binding Domain) and C. elegans nuclear receptor LBD (Ligand Binding Domain) DAF-12. | + | Artificial eukaryotic TF made of Gal4 DBD (DNA Binding Domain) and C. elegans nuclear receptor LBD (Ligand Binding Domain) DAF-12. |
+ | |||
This composite artificial transcription factor will activate any reporter or any gene in general that has a UAS (Upper Activating Sequence) 3' of it's 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. | This composite artificial transcription factor will activate any reporter or any gene in general that has a UAS (Upper Activating Sequence) 3' of it's 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. | ||
[http://partsregistry.org/wiki/index.php?title=Part:BBa_K364328 Go to the official parts registry page for this part] | [http://partsregistry.org/wiki/index.php?title=Part:BBa_K364328 Go to the official parts registry page for this part] | ||
Line 112: | Line 119: | ||
=== [http://partsregistry.org/wiki/index.php?title=Part:BBa_K364320 Gal4-NHR8 Chimeric nuclear receptor] === | === [http://partsregistry.org/wiki/index.php?title=Part:BBa_K364320 Gal4-NHR8 Chimeric nuclear receptor] === | ||
- | Artificial eukaryotic TF made of Gal4 DBD (DNA Binding Domain) and C. elegans orphan nuclear receptor LBD (Ligand Binding Domain). This composite artificial transcription factor will activate any reporter or any gene in general that has a UAS (Upper Activating Sequence) 3' of it's 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. | + | Artificial eukaryotic TF made of Gal4 DBD (DNA Binding Domain) and C. elegans orphan nuclear receptor LBD (Ligand Binding Domain). |
+ | |||
+ | This composite artificial transcription factor will activate any reporter or any gene in general that has a UAS (Upper Activating Sequence) 3' of it's 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. | ||
[http://partsregistry.org/wiki/index.php?title=Part:BBa_K364320 Go to the official parts registry page for this part] | [http://partsregistry.org/wiki/index.php?title=Part:BBa_K364320 Go to the official parts registry page for this part] | ||
=== [http://partsregistry.org/wiki/index.php?title=Part:BBa_K364322 Gal4-NHR23 Chimeric nuclear receptor] === | === [http://partsregistry.org/wiki/index.php?title=Part:BBa_K364322 Gal4-NHR23 Chimeric nuclear receptor] === | ||
Artificial eukaryotic TF made of Gal4 DBD (DNA Binding Domain) and C. elegans orphan nuclear receptor LBD (Ligand Binding Domain). | Artificial eukaryotic TF made of Gal4 DBD (DNA Binding Domain) and C. elegans orphan nuclear receptor LBD (Ligand Binding Domain). | ||
+ | |||
This composite artificial transcription factor will activate any reporter or any gene in general that has a UAS (Upper Activating Sequence) 3' of it's 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. | This composite artificial transcription factor will activate any reporter or any gene in general that has a UAS (Upper Activating Sequence) 3' of it's 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. | ||
[http://partsregistry.org/wiki/index.php?title=Part:BBa_K364322 Go to the official parts registry page for this part] | [http://partsregistry.org/wiki/index.php?title=Part:BBa_K364322 Go to the official parts registry page for this part] | ||
Line 122: | Line 132: | ||
=== [http://partsregistry.org/wiki/index.php?title=Part:BBa_K364323 Gal4-NHR31 Chimeric nuclear receptor] === | === [http://partsregistry.org/wiki/index.php?title=Part:BBa_K364323 Gal4-NHR31 Chimeric nuclear receptor] === | ||
Artificial eukaryotic TF made of Gal4 DBD (DNA Binding Domain) and C. elegans orphan nuclear receptor LBD (Ligand Binding Domain). | Artificial eukaryotic TF made of Gal4 DBD (DNA Binding Domain) and C. elegans orphan nuclear receptor LBD (Ligand Binding Domain). | ||
+ | |||
This composite artificial transcription factor will activate any reporter or any gene in general that has a UAS (Upper Activating Sequence) 3' of it's 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. | This composite artificial transcription factor will activate any reporter or any gene in general that has a UAS (Upper Activating Sequence) 3' of it's 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. | ||
[http://partsregistry.org/wiki/index.php?title=Part:BBa_K364323 Go to the official parts registry page for this part] | [http://partsregistry.org/wiki/index.php?title=Part:BBa_K364323 Go to the official parts registry page for this part] | ||
=== [http://partsregistry.org/wiki/index.php?title=Part:BBa_K364324 Gal4-NHR80 Chimeric nuclear receptor] === | === [http://partsregistry.org/wiki/index.php?title=Part:BBa_K364324 Gal4-NHR80 Chimeric nuclear receptor] === | ||
- | Artificial eukaryotic TF made of Gal4 DBD (DNA Binding Domain) and C. elegans orphan nuclear receptor LBD (Ligand Binding Domain) | + | Artificial eukaryotic TF made of Gal4 DBD (DNA Binding Domain) and C. elegans orphan nuclear receptor LBD (Ligand Binding Domain). |
+ | |||
This composite artificial transcription factor will activate any reporter or any gene in general that has a UAS (Upper Activating Sequence) 3' of it's 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. | This composite artificial transcription factor will activate any reporter or any gene in general that has a UAS (Upper Activating Sequence) 3' of it's 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. | ||
[http://partsregistry.org/wiki/index.php?title=Part:BBa_K364324 Go to the official parts registry page for this part] | [http://partsregistry.org/wiki/index.php?title=Part:BBa_K364324 Go to the official parts registry page for this part] | ||
=== [http://partsregistry.org/wiki/index.php?title=Part:BBa_K364325 Gal4-PXR Chimeric nuclear receptor] === | === [http://partsregistry.org/wiki/index.php?title=Part:BBa_K364325 Gal4-PXR Chimeric nuclear receptor] === | ||
- | Artificial eukaryotic TF made of Gal4 DBD (DNA Binding Domain) and H. sapiens nuclear receptor LBD (Ligand Binding Domain) This composite artificial transcription factor will activate any reporter or any gene in general that has a UAS (Upper Activating Sequence) 3' of it's 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. [http://partsregistry.org/wiki/index.php?title=Part:BBa_K364325 Go to the official parts registry page for this part] | + | Artificial eukaryotic TF made of Gal4 DBD (DNA Binding Domain) and H. sapiens nuclear receptor LBD (Ligand Binding Domain). |
+ | |||
+ | This composite artificial transcription factor will activate any reporter or any gene in general that has a UAS (Upper Activating Sequence) 3' of it's 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. [http://partsregistry.org/wiki/index.php?title=Part:BBa_K364325 Go to the official parts registry page for this part] | ||
=== [http://partsregistry.org/wiki/index.php?title=Part:BBa_K364326 Gal4-EcR Chimeric nuclear receptor] === | === [http://partsregistry.org/wiki/index.php?title=Part:BBa_K364326 Gal4-EcR Chimeric nuclear receptor] === | ||
- | Artificial eukaryotic TF made of Gal4 DBD (DNA Binding Domain) and D. Melanogaster nuclear receptor LBD (Ligand Binding Domain) This composite artificial transcription factor will activate any reporter or any gene in general that has a UAS (Upper Activating Sequence) 3' of it's 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. | + | Artificial eukaryotic TF made of Gal4 DBD (DNA Binding Domain) and D. Melanogaster nuclear receptor LBD (Ligand Binding Domain). |
+ | |||
+ | This composite artificial transcription factor will activate any reporter or any gene in general that has a UAS (Upper Activating Sequence) 3' of it's 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. | ||
[http://partsregistry.org/wiki/index.php?title=Part:BBa_K364326 Go to the official parts registry page for this part] | [http://partsregistry.org/wiki/index.php?title=Part:BBa_K364326 Go to the official parts registry page for this part] | ||
=== [http://partsregistry.org/wiki/index.php?title=Part:BBa_K364327 Gal4-ER Chimeric nuclear receptor] === | === [http://partsregistry.org/wiki/index.php?title=Part:BBa_K364327 Gal4-ER Chimeric nuclear receptor] === | ||
- | Artificial eukaryotic TF made of Gal4 DBD (DNA Binding Domain) and H. Sapiens nuclear receptor LBD (Ligand Binding Domain) This composite artificial transcription factor will activate any reporter or any gene in general that has a UAS (Upper Activating Sequence) 3' of it's 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. | + | Artificial eukaryotic TF made of Gal4 DBD (DNA Binding Domain) and H. Sapiens nuclear receptor LBD (Ligand Binding Domain). |
+ | |||
+ | This composite artificial transcription factor will activate any reporter or any gene in general that has a UAS (Upper Activating Sequence) 3' of it's 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. | ||
[http://partsregistry.org/wiki/index.php?title=Part:BBa_K364327 Go to the official parts registry page for this part] | [http://partsregistry.org/wiki/index.php?title=Part:BBa_K364327 Go to the official parts registry page for this part] | ||
=== [http://partsregistry.org/wiki/index.php?title=Part:BBa_K364329 ER DBD-VDR hinge-ER LBD Chimeric nuclear receptor] === | === [http://partsregistry.org/wiki/index.php?title=Part:BBa_K364329 ER DBD-VDR hinge-ER LBD Chimeric nuclear receptor] === | ||
- | Artificial eukaryotic TF made of Estrogen receptor's DBD (DNA Binding Domain), a vitamin D3 receptor hinge domain and H. sapiens nuclear receptor LBD (Ligand Binding Domain) This composite artificial transcription factor will activate any reporter or any gene in general that has a UAS (Upper Activating Sequence) 3' of it's 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. | + | Artificial eukaryotic TF made of Estrogen receptor's DBD (DNA Binding Domain), a vitamin D3 receptor hinge domain and H. sapiens nuclear receptor LBD (Ligand Binding Domain). |
+ | |||
+ | This composite artificial transcription factor will activate any reporter or any gene in general that has a UAS (Upper Activating Sequence) 3' of it's 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. | ||
[http://partsregistry.org/wiki/index.php?title=Part:BBa_K364329 Go to the official parts registry page for this part] | [http://partsregistry.org/wiki/index.php?title=Part:BBa_K364329 Go to the official parts registry page for this part] | ||
+ | |||
+ | <html><br><a name="composite"></a><br></html> | ||
== Eukaryotic expression vectors - Composite Parts == | == Eukaryotic expression vectors - Composite Parts == | ||
Line 154: | Line 176: | ||
=== [http://partsregistry.org/wiki/index.php?title=Part:BBa_K364332 TRE-Gal4-PXR-PolyA in PSB1A3 (expression vector)] === | === [http://partsregistry.org/wiki/index.php?title=Part:BBa_K364332 TRE-Gal4-PXR-PolyA in PSB1A3 (expression vector)] === | ||
- | Expression vector from standard BioBrick parts, expressing Gal4 - human Pregnane X receptor LBD composite part | + | Expression vector from standard BioBrick parts, expressing Gal4 - human Pregnane X receptor LBD composite part. |
+ | |||
+ | The Gal4-PXR Chimeric nuclear receptor is an artificial eukaryotic TF made of Gal4 DBD (DNA Binding Domain) and H. sapiens nuclear hormone receptor LBD (Ligand Binding Domain) which can be expressed by this expression vector. | ||
+ | |||
+ | The minimal CMV promoter ensures a continuous expression of the chimeric construct, but furthermore it can be expressed in an inducible form using the Tetracyclin-controlled transcriptional activation system. | ||
[http://partsregistry.org/wiki/index.php?title=Part:BBa_K364332 Go to the official parts registry page for this part] | [http://partsregistry.org/wiki/index.php?title=Part:BBa_K364332 Go to the official parts registry page for this part] | ||
=== [http://partsregistry.org/wiki/index.php?title=Part:BBa_K364333 TRE-Gal4-EcR-PolyA in PSB1A3 (expression vector)]=== | === [http://partsregistry.org/wiki/index.php?title=Part:BBa_K364333 TRE-Gal4-EcR-PolyA in PSB1A3 (expression vector)]=== | ||
- | Expression vector from standard BioBrick parts, expressing Gal4 - Ecdysone receptor LBD composite part. | + | Expression vector from standard BioBrick parts, expressing Gal4 - Ecdysone receptor LBD composite part. |
+ | |||
+ | The Gal4-EcR Chimeric nuclear receptor is an artificial eukaryotic TF made of Gal4 DBD (DNA Binding Domain) and D. Melanogaster nuclear hormone receptor LBD (Ligand Binding Domain) which can be expressed by this expression vector. | ||
+ | |||
+ | The minimal CMV promoter ensures a continuous expression of the chimeric construct, but furthermore it can be expressed in an inducible form using the Tetracyclin-controlled transcriptional activation system. | ||
[http://partsregistry.org/wiki/index.php?title=Part:BBa_K364333 Go to the official parts registry page for this part] | [http://partsregistry.org/wiki/index.php?title=Part:BBa_K364333 Go to the official parts registry page for this part] | ||
Latest revision as of 09:39, 26 October 2010
|