I cloned human ACSL1 promoter (2.3 kb) in pGL3 basic vector and when I did the luciferase assay. My ACSL1 promoter basal activity is similar to pGL3 basic-luc, Generally promoter of your gene of interest always shows higher basal promoter activity than pGL3 basic control vector. I did sequencing of pGL3-hACSL1 promoter contract- sequencing result is ok.
Does anyone have any idea what happened?
Thanks,
Amar
Hi Nilabh,
Thanks for your suggestions,
I didn't use GFP but I used renilla as a internal control and also 0.5% FBS. well actually promoter is active, when i treated with ligand prmoter activity was increased several folds, only problem is basal activity of promoter is similar like pGL3 basic luc
Is the promoter situated in the right position/direction? are you sure you are using the right clone?
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