Figure 4

Packaging of hY RNAs requires the NC zinc finger domains. HeLa cells were co-transfected with pcDNA-APO3G-MycHis together with pNL4-3ΔVif (43ΔVif) or pDB653ΔVif. Viruses were harvested 24 h after transfection and purified as described in Methods. (A) Virus production and packaging of APO3G was monitored by immunoblot analysis using an aliquot of the purified, concentrated virus preparations. APO3G encapsidation was identified using a polyclonal APO3G-specific peptide antibody. Viral capsid proteins (CA) were identified using an HIV-positive human patient serum (APS). (B) RNAs were extracted from purified, concentrated viruses and amplified by RT-PCR using primer pairs specific for HIV-1 RNA or host RNAs as indicated on the left and detailed in table 1. RNA extracted from C-HelpΔVif preparations in figure 3 was included as control. RT-PCR products were separated on 1% agarose gels and visualized by staining with ethidium bromide.