FIG. 2.
Analysis of the products of the 6-mer and 9-mer RT-PCRs of the US11 RNA pull down. (A) Fractionation of the products of the 6-mer RT-PCR on an ethidium bromide-stained agarose gel. Lanes: 1, 1-kb ladder marker; 2, RT-PCR with RNA isolated using GST alone in the pull down; 3, RT-PCR with RNA isolated using GST-US11 fusion protein in the pull down (the arrowhead indicates the band that hybridizes to the Δ34 probe in the blot in panel B); 4, 100-bp marker. (B) Southern blot of the gel shown in panel A hybridized with an in vitro-transcribed 32P-labeled Δ34 probe. (C) Fractionation of the products of the 9-mer RT-PCR on an ethidium bromide-stained agarose gel. Lanes: 1, 1-kb ladder marker; 2, control RT-PCR using Δ34-specific primers and RNA isolated using GST alone in the pull down; 3, control RT-PCR using Δ34-specific primers and RNA isolated using GST-US11 fusion protein in the pull down; 4, RT-PCR with random and anchor primers and RNA isolated using GST alone in the pull down; 5, RT-PCR with random and anchor primers and RNA isolated using GST-US11 fusion protein in the pull down. (D) Diagram of the three coterminal transcripts UL12, UL13, and UL14 showing the position of the US11-pulled-out 12/14 RNA and the sequence of the 585-nt 12/14 RNA. Thick lines, coding regions; thin lines, 5′ and 3′ UTRs; (A)n, end of the message.