I want to immortalize cells using hTERT and SV40 using vectors:
pLXSN/SV40, the SV40 would be expressed from 5`LTR promoter, the plasmid
gives resistance for neomycin,
pMSCVhyg/hTERT, the hTERT would be also expressed from 5`LTR promoter, the plasmid gives resistance for hygromycin.
is anyone aware if the two genes maybe expressed form the same promoter (5`LTR) and still work. Wont it cause any promoter interference?
Divaker Choubey
Assuming that both plasmid DNAs are of comparable high quality (free from any associated proteins) and transfected in certain ratio (1:1 or 1:2), it should be possible to select cells for both drug resistance. As controls, cells without any plasmid DNA transfection and singly transfected plasmd DNA should be included.
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