The pRL-CMV Vector(a,b,c) (Figure 1) is intended for use as an internal control reporter and may be used in combination with any experimental reporter vector to co-transfect mammalian cells. All of Promega’s pRL Reporter Vectors contain a cDNA (Rluc)(a) encoding Renilla luciferase, which was originally cloned from the marine organism Renilla reniformis (sea pansy; 1). As described below, the Renilla luciferase cDNA contained within the pRL Vectors has been modified slightly to provide greater utility.

The pRL-CMV Vector contains the CMV(b) enhancer and early promoter elements to provide high-level expression of Renilla luciferase in co-transfected mammalian cells. Renilla luciferase is a 36kDa monomeric protein that does not require posttranslational 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,e) Technical Manual (#TM040).

To avoid DNA methylation, all pRL Vectors are isolated from a dam-/dcm- E. coli K host strain. Therefore, initial propagation of the pRL-CMV Vector should be conducted in an appropriate E. coli host strain lacking endogenous restriction endonuclease activity (e.g., JM109).