Figure 5

PCR analysis of genomic DNA from NIH3T3 cells transfected with semi-replicative retroviral vectors. (A) The generation of full-length MLV genomes was analyzed by PCR using the primers p1 and p2 (see Fig. 4). Full-length MLV generates an 800 bp PCR fragment, semi-replicative retroviral vectors should not give rise to a DNA fragment because the primers do not bind to the same genome. DNA was transfected in different molar ratios as indicated. The first number indicates the molar ratio of the gag/pol plasmid and the second the Env encoding plasmid.(B) The stability of the GFP sequences inserted into the Env gene was analyzed by PCR using the primers p3 and p4 (see Fig. 4). The gfp-env sequence gives rise to a 1.5 kb fragment and wt env to an 800 bp fragment. Untransfected NIH3T3 cells were cultured in parallel and analyzed identically. The data are given as negative at days 13 and 32. NTC, no template control.