I have a 2.5kb insert including fluorescent proteins. The problem I am having is that I see nice expression of these from a Topo His plasmid in 293 cells, how ever when I clone this segment into mscv hygro I see no expression in 293 cells. I have done restriction digests on the plasmid to check for presence and orientation of my insert and sequenced, all looks fine. Simultaneously i ligated only GFP into the same plasmid and I see expression of GFP no problems.
expression in TopoTA but not in mscv
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