Stable transfection problem - About co-transfection (Feb/05/2007 )

Dear all,

I am going to establish a stable HepG2 cell line harboring a promoter+luciferase construct. I co-transfect the cell with the promoter+luciferase plasmid (pGL3) and pcDNA3.1. I just wonder whether I can use a pcDNA3.1 inserted with EGFP instead of empty pcDNA3.1 because I can use the fluorescence signal to look for stable transfectants and I can check the luciferase activity of those fluorescence emitting clones. Will the luciferase activity be affected by the presence of EGFP protein inside the clones?

Please advice.

Thanks

Charles

-charles_mozart-

QUOTE (charles_mozart @ Feb 6 2007, 04:27 AM)

that is an interesting task; although I did not check if luciferase activity is affected by EGFP, I suppose it should not; there will only difficulties if you like to measure EGFP emission and luminescence in parallel but for sequential measurement there seem to be theoretically no problem; the luciferase signal in co-transfected cells (EGFP and luciferase) may be higher as in single luciferase transfected cells as luminescence may excite EGFP fluorescence signal which will contribute to the total luminescence signal

-The Bearer-

Just be sure that EGFP can be harmful to cells. You already have neo resistant gene in pcDNA for selection. EGFP may not add any more value to it.

-genehunter-1-