The pRL-SV40 Vector(a,b) (Figure 1) is intended for use as an internal control reporter and may be used in combination with any experimental reporter vector to cotransfect mammalian cells. All of Promega’s pRL Reporter Vectors contain a cDNA (Rluc) encoding Renilla luciferase, which was originally cloned from the marine organism Renilla reniformis (sea pansy; 1). As described below, the Renilla luciferase cDNA(b) contained within the pRL Vectors has been modified slightly to provide greater utility.
The pRL-SV40 Vector contains the SV40 enhancer and early promoter elements to provide high-level expression of Renilla luciferase in cotransfected mammalian cells. Renilla luciferase is a 36kDa monomeric protein that does not require post-translational modification for activity (2). Therefore, like firefly luciferase, the enzyme may function as a genetic reporter immediately following translation. For information about the use of this plasmid in conjunction with a reporter vector containing the firefly luciferase gene, refer to the Dual-Luciferase ® Reporter Assay System (c,d) Technical Manual (#TM040).
The pRL Vectors are isolated from a dam–/dcm– E. coli K host strain, allowing digestion with restriction enzymes that are sensitive to dam and dcm methylation.
The GenBank®/EMBL Accession Number for the pRL-SV40 Vector is AF025845.