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. 2006 Feb;80(3):1242–1249. doi: 10.1128/JVI.80.3.1242-1249.2006

FIG. 2.

FIG. 2.

Analyses of the protein and RNA contents of cell-free virions and viral DNA generated by infection. (A) Western analysis of the cell-free virion proteins. Cell-free virions were harvested 2 days after transfection and examined by Western blotting using rabbit anti-MLV MA and CA antibodies. (B) Comparison of the amounts of MLV vector RNA packaged in cell-free virions. RNAs isolated from virions with similar levels of RT activity were used in the real-time RT PCR analysis; primers and probes in hygro were used to detect SR2-2GFP vector RNA. The amount of SR2-2GFP RNA packaged in pLGPS-derived virions was set to 100% in each experiment. Data from three independent experiments are shown as means ± standard errors. (C) Synthesis of vector DNA upon infection. Quantitative real-time PCR analysis of the early (RU5; left panel) and late (U5Ψ; right panel) viral DNA products generated by viral infection. Viral DNAs from D17 cells infected by viral stocks with similar levels of RT activity 6 h and 12 h postinfection were used in the real-time PCR analysis. RU5 and U5Ψ copy numbers synthesized by pLGPS-derived virions were set to 100% in each experiment. Data from five independent sets of experiments are shown as means ± standard deviations. WT, pLGPS; Δ12, pMLVCAΔ12; Δ12/PR−, pMLVCAΔ12/PR−; PR−, pMLVPR−; mock, mock transfection.