Team:ECNUAS/Composite Parts
Favorite Composite Part: BBa_K3704001
Biology
Composite part BBa_K3704001 consists of coding sequence of firefly luciferase under control of glucocorticoid response element (GRE). Luciferase can catalyze the oxidation of luciferin to oxyluciferin. During the oxidation of luciferin, it will emit bioluminescence. Glucocorticoid receptor (GR) belongs to the steroid hormone receptor subclass of nuclear receptors, which controls physiological processes by activating and inhibiting specific target genes (PMID: 15379669). The ligand-activated receptor dimer activates gene expression by binding to GRE in the promoter region of the glucocorticoid regulatory gene. BBa_K3704001 forms a reporting system which are regulated by GR agonist (increases the fluorescence value) and GR antagonist (reduces the fluorescence value that is stimulated by GR agonist).
Functional verification of the GRE-Fluc reporting system
In order to test the function of the GRE-Fluc reporting system, GRE-Fluc containing plasmid and another two plasmids which produce GR (part BBa_K3704002) and Renilla luciferase (BBa_K3522012, used as an internal reference gene to provide a unified baseline for experiments), respectively, were co-transfected into HEK293T cells. With addition of GR agonist (Dexamethasone, Dex) into the cell culture, significant value of luciferase activity was detected (Figure 1). While, in the presence of Dex and a GR antagonist (Mifepristone, Mife), the luciferase activity reduced to a value similar to the blank control (DMSO).
Figure 1. Relative luciferase activity in the presence of GR agonist (Dexamethasone, Dex) and GR antagonist (Mifepristone, Mife).
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