I transformed by BCG culture with my gene of interest, and after growth I set up an experiment with a drug that is used as a control for the expression of my gene: if my gene is overexpressed, resistance to this drug should be observed, but unfortunately, there are no differences between my control BCG and my over expresser. I need a brainstorm of ideas that could explain this results!
What plasmid are you expressing your gene in? You need to know the optimal concentration of you drug that kills your non transfected cells and selects for your gene expression.
For example- you're expressing pEGFP which has a G418 selection cassette- an antibiotic. I would perform a kill curve- which means I would plate out the cells I'm planning to express my pEGFP construct in, at 50% confluence into a 6 well dish. I would and treat them with a range of G418 concentrations.
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