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. 2002 Sep;76(18):9096–9102. doi: 10.1128/JVI.76.18.9096-9102.2002

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Copyright © 2002, American Society for Microbiology

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FIG. 3. — Alteration of tRNAin protease-negative HIV-1 as a result of overexpression of tRNA. Protease-negative viruses were produced from COS7 cells transfected with BH10.P(−) or BH10.P(−)Lys3. (A) 2D-PAGE analysis of low-molecular-weight viral RNA. Total viral RNA was 3′ end labeled with [³²P]pCp and then electrophoresed. Conditions for 2D-PAGE and labeling of spots is as described in Fig. 2A. (B) Western blots of viral lysates, probed with anti-CA and anti-RT. The results, quantitated by phosphorimaging, are listed in panel C as the GagPol/Gag ratios. (C) Incorporation of tRNA^Lys into HIV-1. Dot blots of viral RNA were hybridized with DNA probes complementary to tRNAalone, to tRNA^Lys (both tRNAand tRNA), and to viral genomic RNA. The results were quantitated by phosphorimaging, and the tRNA/genomic RNA or tRNA^Lys/genomic RNA ratios are listed in panel C.

Inline graphic — Alteration of tRNAin protease-negative HIV-1 as a result of overexpression of tRNA. Protease-negative viruses were produced from COS7 cells transfected with BH10.P(−) or BH10.P(−)Lys3. (A) 2D-PAGE analysis of low-molecular-weight viral RNA. Total viral RNA was 3′ end labeled with [³²P]pCp and then electrophoresed. Conditions for 2D-PAGE and labeling of spots is as described in Fig. 2A. (B) Western blots of viral lysates, probed with anti-CA and anti-RT. The results, quantitated by phosphorimaging, are listed in panel C as the GagPol/Gag ratios. (C) Incorporation of tRNA^Lys into HIV-1. Dot blots of viral RNA were hybridized with DNA probes complementary to tRNAalone, to tRNA^Lys (both tRNAand tRNA), and to viral genomic RNA. The results were quantitated by phosphorimaging, and the tRNA/genomic RNA or tRNA^Lys/genomic RNA ratios are listed in panel C.