RNA from the pHyVec7 transgene is trans-spliced.

<p>RNA from the operon transgenic line was reverse-transcribed to produce first strand cDNA. The cDNA was amplified with primers for the spliced leader SL-B1 and the DsRed2 coding sequence. The top line shows sequence between the RFC140 and <i>flp</i> genes extending from the polypyrimidine tract associated with the trans-splicing AG acceptor dinucleotide (doubly underlined) to the start codon of the <i>flp</i> gene (shaded in gray). The second line shows the sequence from a previously identified <i>flp</i> EST containing the SL-B1 spliced leader sequence <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0031643#pone.0031643-Steele1" target="_blank">[13]</a>; the spliced leader sequence is highlighted in yellow, and the <i>flp</i> start codon is shaded. The third line shows sequence from the RT-PCR product obtained from the pHyVec7 transgenic line using SL-B1 and DsRed2 primers. The sequence corresponding to the SL-B1 primer is highlighted in green, and the DsRed2 start codon is shaded.