Figure 3

Human ORFs with divergent sequences are not expressed in testis. (A) Diagram of the RT-PCR approach used to distinguish expression of transcripts. Black arrows denote primer sets used to amplify both parent gene and retroposed sequences. The fraction next to each retroposed sequence shows the number of unique nucleotide residues in the amplified product. (B) TPI1-rs1, PGAM1- rs7, and ENO1- rs1 transcripts were not detected in pooled human testis RNA samples with RT-PCR using primers that amplify both the retroposed sequence and the parent glycolytic enzyme, followed by single-strand conformation polymorphism (SSCP) gel electrophoresis. PCR products amplified from human genomic DNA (G1 and G2; two individuals) show the expected position of transcripts from retroposed sequences. PCR products amplified from pooled testis total RNA are shown in lanes T1, T2, and T3 (triplicate cDNA preparations).