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Figure 3 | Retrovirology

Figure 3

From: The prototype foamy virus protease is active independently of the integrase domain

Figure 3

Analysis of virus infectivity and PR activity in vivo using different GagPol-fusion proteins. (A) A schematic diagram of the GagPol precursor protein is depicted on top of the panels: CS, PR cleavage site. (B) Comparison of the virus titer of FV codon optimized vector system (VS) [11] or the GagPol fusion plasmid cotransfected with an env expression construct and pMD9 as gfp-encoding genome. Additional amounts of a gag expression plasmid (pcoPG4) used for cotransfections are indicated. The transfected DNA amounts are indicated in [μg]. Transfections were performed in triplicate assays. The error bars represent the standard deviation. (C) Western blotting analysis of the samples from Figure 3A. The cellular and viral Gag/Pol amounts and processing using Gag and RT specific antibodies were analysed (VS: FV vector system [11]). (D) The virus release of GagPol fusion is Env dependent but genome independent and the processing of Gag is IN independent. Western blotting analysis of the cellular and viral Gag/Pol amounts and processing. HEK 293 T (2×105) cells were transfected with pcoPE, pMD9, pcoPG4, and either pGagPol or pGagPR-RT. The transfected DNA amounts are indicated in [μg]. Analysis of the GAPDH expression served as a loading control.

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