Evaluation of a novel luciferase reporter construct: a positive control plasmid for reporter gene assay

  • T Tencomnao
  • V Rakkhitawatthana
  • K Sukhontasing
Keywords: Reporter gene plasmid, luciferase assay, cytomegalovirus promoter/enhancer, human melanoma cell line.

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 cisacting 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.
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eISSN: 1684-5315