Figure 4

Detection of T1–T2 chimeric transcripts in the testis. (A) RT-PCR profiling of T1–T2 chimeric transcripts. Two rounds of nested PCR were performed using oligo(dT)-primed RT products of the testis mRNA. In the first-round PCR, the consensus Ex1a-B and the T1- or T2-specific 3'-UTR primers, T1SP-R1 and T2SP-R1, respectively, were used (see Figure 1A for relative positions and Table 2 for sequence details); in the second-round PCR, the exon 3 (T1) or exon 2 (T2)-based T2SP-F1 or T1SP-F1 primers were used in combination with T1SP-R1 and T2SP-R1. The T1 and T2 plasmids were used as controls in both rounds of PCR (lanes 1–2 and 4–5). The PCR products (with designations in brackets) generated in the second-round PCR (lane 6) were cloned and sequenced. (B) Schematic depiction of the PCR products derived from (A). The exons of the transcripts are shown using the same colour code and exon designation as in Figure 2. Primers used in the second-round PCR are also shown.