Fig. 2.

One-step RT-PCR analysis of transcripts at the ORF 33 locus. A) Diagram of ORFs and transcripts at the ORF 33 locus (after (Detvisitsakun et al., 2005). The viral ubiquitin gene (ubi), encoded on the opposite strand with transcripts originating within ORF34, is also included in the diagram. Solid arrows are previously mapped transcripts and dashed lines indicate an observed transcript whose 5’ and 3’ ends have not been mapped (3.1 Kb) and a putative ORF 34 transcript (X). The solid line (*) represents a polycistronic message confirmed by one-step RT-PCR. The locations of primers used for analysis are shown. B) Transcripts through each ORF at 48 h p.i. with vTRPV4 following transfection with ds33 or dsCAT. ORFs, primer pairs and dsRNAs are indicated. C) Transcripts across adjacent ORFs as indicated at the top of the panel. Primer pairs and dsRNAs are indicated. 28S RNA was used as a control. Black box in C and * in A and C indicate a transcript that includes FGF, ORF33 and ORF34. A non-specific band (~ 750 bp) was detected when F1 primer was used (arrow, in B and C).