Difference between revisions of "Part:BBa K364320"

 
 
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<partinfo>BBa_K364320 short</partinfo>
 
<partinfo>BBa_K364320 short</partinfo>
  
 
Gal4 DBD - NHR 8 C. elagans LBD
 
Gal4 DBD - NHR 8 C. elagans LBD
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Artificial eukaryotic TF made of Gal4 DBD (DNA Binding Domain) and C. elegans orphan nuclear receptor LBD (Ligand Binding Domain)
 +
 +
NHR-8 LBD
 +
 +
Nuclear hormone receptor family member nhr-8.
 +
 +
The nhr-8 gene encodes a nuclear hormone receptor homolog; nhr-8 (ok186) mutants have abnormally low resistance to the toxins colchicine and chloroquine. NHR-8 functions in the nematode xenobiotic defense system.
 +
 +
Gal4 DBD
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 +
This protein is a positive regulator for the gene expression of the galactose-induced genes such as GAL1, GAL2, GAL7, GAL10, and MEL1 which encode for the enzymes used to convert galactose to glucose. This protein contains a fungal Zn(2)-Cys(6) binuclear cluster 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.
 +
 +
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. [http://2010.igem.org/Team:Debrecen-Hungary]
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[[Image:pSB1C3-Gal4-NHR8.gif|800px|thumb|center|Picture of gel electrophoresis: Gal4-NHR8 in pSB1C3 resulting an insert of 953 bp. Circular pSB1C3-RFP, pSB1C3-Gal4-NHR8 and restricted PMAT-NHR8 can be seen as control on the gel.]]
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[[Image:nhr64.jpg|center|600px|thumb|Activation of Gal-NHR8 and Gal-NHR64 to oil sand extractions.]]
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[[Image:nhr8.jpg|center|300px|thumb|Activation of NHR8 to Blesse bleu cheese extract.]]
  
 
<!-- Add more about the biology of this part here
 
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Latest revision as of 07:45, 21 September 2011

Gal4-NHR8

Gal4 DBD - NHR 8 C. elagans LBD

Artificial eukaryotic TF made of Gal4 DBD (DNA Binding Domain) and C. elegans orphan nuclear receptor LBD (Ligand Binding Domain)

NHR-8 LBD

Nuclear hormone receptor family member nhr-8.

The nhr-8 gene encodes a nuclear hormone receptor homolog; nhr-8 (ok186) mutants have abnormally low resistance to the toxins colchicine and chloroquine. NHR-8 functions in the nematode xenobiotic defense system.

Gal4 DBD

This protein is a positive regulator for the gene expression of the galactose-induced genes such as GAL1, GAL2, GAL7, GAL10, and MEL1 which encode for the enzymes used to convert galactose to glucose. This protein contains a fungal Zn(2)-Cys(6) binuclear cluster 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.

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. [http://2010.igem.org/Team:Debrecen-Hungary]

Picture of gel electrophoresis: Gal4-NHR8 in pSB1C3 resulting an insert of 953 bp. Circular pSB1C3-RFP, pSB1C3-Gal4-NHR8 and restricted PMAT-NHR8 can be seen as control on the gel.
Activation of Gal-NHR8 and Gal-NHR64 to oil sand extractions.
Activation of NHR8 to Blesse bleu cheese extract.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 218
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 137
    Illegal SapI.rc site found at 545