Abstract

Reporter gene technology has been increasingly important in the post-genomic era to explain human complexity and diversity. The pGL3-Basic vector has been prevalently used as a tool for analyzing cis-acting elements critical for transcriptional mechanisms. In this work, we constructed and evaluated the pGL3-Basic plasmid containing the cytomegalovirus (CMV) enhancer/promoter aiming to establish a positive control of pGL3-Basic vector. Using a human melanoma cell line UACC-903 for transient transfection, the novel luciferase reporter construct, pGL3-CMV, showed an extremely high transcriptional activity approximately 4,260-fold greater than that of pGL3-Basic, indicating its qualification as a positive control for luciferase reporter gene assays.

Key words: Reporter gene plasmid, luciferase assay, cytomegalovirus promoter/enhancer, human melanoma cell line

Abbreviation

bp, base pair; CAT, chloramphenicol acetyltransferase; CMV,cytomegalovirus; FBS, fetal bovine serum; GFP, green fluorescent protein; PCR, polymerase chain reaction; RPMI, Rosewell Park Memorial Institute; and SV40,simian virus 40.