We've been trying to express GFP in mammalian cell lines with very little
success. We've used both vectors from Clonetech (pGFP-N1) and a vector
constructed here with comparable lack of success. The cell lines tried were
HeLa, 293, NRKE, all with no success. A different collegue has said that he
sees expression in MCF7 cells, but I haven't tried these yet. I've heard
scuttlebutt about these sorts of problems elsewhere. Does anyone have any
feel for what (if any) are the barriers to expression in mammalian cells?
I've heard that codon usage may be a problem. Any experience with this? We
haven't done Southerns and Northerns yet, but I have no reason to think that
these vectors won't fire in the cell types listed above. Any suggestions
would be greatly appreciated.
SK at LILLY.COM