pCAG-GFP plasmid contains a chimeric intron and GFP, I cut this plasmid with EcoR I and Not I , so the GFP sequence was removed , then I insert my interested protein(95kd) gene to replace GPF. Sequencing result showed that transfected this construct to mammalian cell to overexpress my gene. After western blotting detect the proteins, there are two bands in overexpressed cell, one is the same size with ednogenous protein, the other is 10kd 's bigger than my protein. I can not understand why did the bigger protein produced, since there were no tag after remove GFP.
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