Renilla Luciferase DNA (Promega) without (pMB 341) or with (pMB 725, a gift of Joe Avruch) an IRES sequence was transcribed in vitro. Capping reactions were conducted with the ScriptCap m7G capping system as described (CellScript). IRES-containing RNA was not capped. RNA was incubated at 10 nM in extracts at 20°C for 1 hr., and luciferase activity measured using the Dual-Glo Lucifearse Assay System (Promega).
Renilla luciferase dna
Manufactured by Promega
Sourced in United States
Renilla Luciferase DNA is a genetic reporter system that produces bioluminescence when Renilla luciferase enzyme is expressed in cells. It can be used to measure gene expression or promoter activity.
Automatically generated - may contain errors
2 protocols using renilla luciferase dna
1
In Vitro Transcription and Luciferase Assay
2
DEX-responsive Luciferase Assay in ST2 Cells
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An expression vector containing the luciferase-reporter gene under the control of a DEX-responsive element (pBV2-MMTV-LUC) was gifted by Drs H Noguchi and I Abe of the University of Shizuoka (Shizuoka, Japan).50 (link) ST2 cells (RIKEN BioResource Center) were seeded at a density of 2.5×104 cells/well on Ti plates (10×10×1 mm) in 24-well plates. ST2 cells were maintained in RPMI 1640 supplemented with 10% FBS, 100 µg/mL penicillin, and 100 U/mL streptomycin in continuous culture at 37°C under a humidified atmosphere with 5% CO2 and 95% air. After incubating for 1 day, cells were co-transfected with pBV2-MMTV-LUC and thymidine kinase promoter-driven Renilla luciferase DNA (Promega Corporation, Fitchburg, WI, USA) using FuGENE6 (Promega). After an additional 24 h, luciferase activity was measured using the dual-luciferase reporter assay system (Promega) and an AutoLumat LB953 luminometer (Berthold Technologies, Oak Ridge, TN, USA) according to the manufacturer’s instructions.
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