Hiyoshi Corporation Provides CALUX®Assay
  Mechanism of CALUX Cell   Accuracy   Analysis Process
  HOMEMethodMechanism of CALUX Cell
 
The recombinant cell line used in this assay (H1L6.1c2) was generated by stably transfecting the plasmid pGudLuc6.1 into mouse hepatoma (Hepa1c1c7) cells. The pGudLuc6.1 plasmid contains the CYP1A1 dioxin-responsive domain (inclusive of four DREs) upstream of the firefly luciferase gene.
 
 
Halogenated aromatic hydrocarbons (HAH), including dioxins bind to an intracellular receptor called the aryl hydrocarbon receptor (Ah Receptor) and activate the receptor.
The HAH-Ah Receptor complex then travels into the nucleus of the cell.
Activated HAH-Ah Receptor then binds to specific sequences in the DNA called dioxin responsive elements (DRE).
The binding of the HAH-Ah Receptor complex to the XRE induces the expression of XRE-associated genes.
Luciferase gene expression is increased.
The induced messenger RNA (mRNA) is released into the cytoplasm.
The mRNAs are translated into new proteins.
New proteins produce the biological and toxicological effects that are observed following exposure to dioxin and related HAHs.
A Dioxin TEQ is determined from a dose response curve of the luminescence produce by the enzymatic activity of the induced luciferase reporter protein.


 

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