The cassettes containing the PTGS2 wild-type 3'-UTR (rs5275 T), or its variant (rs5275 C), were subcloned into PR constructs previously generated in XbaI sites, so that the regulatory regions (PR and 3'-UTR) of PTGS2 would flank the luciferase
acts on a molecule called luciferin, causing it to emit light.
Previous efforts to create light-emitting plants have relied on genetically engineering plants to express the gene for luciferase
, but this is a laborious process that yields extremely dim light.
The team used enzyme called luciferase
, which is the same substance that gives fireflies their amazing flashing behinds at night.
KeratinoSens[TM] cells are derived from the human keratinocyte cell line HaCat, containing a luciferase
gene that is under the control of a constitutive promoter fused with the antioxidant response element (ARE) from a gene that is known to be up-regulated by contact sensitizers.
For the pgCAR dual-luciferase
assay, NR1 as the CAR responsive element was cloned upstream of the firefly luciferase
reporter gene to pick up the ligands which regulate pgCAR causing altered reporter gene expression.
reporter assays confirmed that miR-106b-5p binds to PTEN mRNA 3' untranslated region site.
They have developed a complementary strategy based on capturing receptor-proximal conformational profiles using intra-molecular BRET-based sensors composed of a Renilla luciferase
donor engineered into the carboxy-terminus and CCPGCC motifs which bind fluorescent hairpin arsenical dyes engineered into different positions in the AT1 angiotensin II receptor and the PGF2?
and its exogenously supplied substrate, luciferin, can be used to measure ATP because the chemiluminescent reaction is ATP-dependent.
Further, MGDG treatments significantly inhibited luciferase
activities of constructs containing liver X receptor response element in FASN promoter region without altering the levels of mRNA encoding transcription factors.
single-strand annealing (SSA) reporter assay was performed as described previously (Wang et al.
Blumberg, University of California, Irvine], 80 ng UASx4 TK-luc [contains four copies of the GAL4 upstream activating sequence and the herpes virus thymidine kinase promoter (-105/+51) driving firefly luciferase
] and 4 ng of pRL vector (encoding Renilla luciferase
to control for transfection efficiency).