Fig. 5.

3′-RACE analysis of the AL3 transcript. (a) Schematic representation of the AL3 ORF, primers used in 3′ RACE (Supplementary Table S1) and positions of putative polyA addition signals. The position of the stop codon of the AL3 ORF is indicated by +856. (b) Total RNA was prepared from RS cells infected with the designated virus (R, dLAT2903R; 2903, dLAT2903), DC-LAT6 cells or TG of latently infected mice (C57/BL/6 mice). 3′-RACE was performed according to the manufacturer's recommendations (Ambion). Specific primers used for PCR and nested PCR are shown. Lane M is a 100 bp DNA Ladder (Invitrogen). (c) Summary of AL3 3′-RACE results. RT-PCR products were cloned and sequenced. AL3 indicates DNA sequences that correspond to the AL3 cDNA sequence. Dashes in the LAT sequence indicate identity to the AL3 sequence. The position of the AL3 3-for primer used for mapping the 3′ terminus of the AL3 transcript is indicated. Arrows denote the 3′ terminus of the AL3 transcript. Images were prepared from gels as described in Fig. 2.