Expression of Gluc-Fluc dual luciferase plasmid after transfection into MB49 bladder cells / 南方医科大学学报

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the expression characteristics of Gluc-Fluc dual luciferase plasmid after its transfection into MB49 bladder cells.</p><p><b>METHODS</b>pAAV2neoCAG-Gluc-2A-Fluc and pAAV2neo-Gluc plasmids were separately transfected into MB49 cells via LipofectamineTM2000. The Gluc activity in the cell culture supernatant and the Fluc activity in the cells were detected by luminometer and Lumina Imaging system.</p><p><b>RESULTS</b>The luminometer result showed that the activity of Gluc in the supernatant increased gradually in a cell number- and time-dependent manner, while Fluc activity in the cells increased with the cell number but not with time. The Lumina Imaging system showed that Gluc-Fluc was successfully expressed in MB49 bladder cells and cell lines with stable Gluc-Fluc expression were obtained after G418 selection.</p><p><b>CONCLUSION</b>Gluc in the dual luciferase plasmid retains its expression characteristics. Due to the advantages of Fluc in localization in living imaging and the easy quantitative detection of Gluc, the dual luciferase plasmid, after transfection in MB49 bladder cells, allows reliable and dynamic detection of tumor growth in animal models.</p>