Cell lines selected with G418 but no overexpression of GOI
Posted 09 July 2013 - 12:03 PM
I transfected cells with empty vector and one containing my GOI. I selected using G418. However when I do the IB for my GOI there is no expression. The GOI is under a CMV promoter, while the Neomycin is under a SV40 promoter. I was wondering if anyone has had a similar experience. Could it be that overexpression of the GOI is toxic to cells and therefore they are no longer expressing from the CMV promoter? I know the CMV promoter is active in these cells as transient transfections have resulted in ample overexpression of the protein. Any insights would be greatly appreciated.
Posted 09 July 2013 - 12:29 PM
Posted 09 July 2013 - 12:32 PM
Posted 09 July 2013 - 12:35 PM