Figure 2.

RT–PCR and protein expression analysis of the bank vole HBA-T3 globin gene. (a) Genomic structure of the gene. Alleles containing PTCs in exons 2 and 3, referred to as, respectively, the B allele and C allele, are aligned with a non-PTC allele referred to as the A allele. (b) RT–PCR and PCR assays followed by gel electrophoresis were performed to assess the amplification of HBA-T3 cDNA and genomic DNA (gDNA) using paralogue-specific primers spanning the entire coding region. Data represent HBA-T3 cDNA amplification from mRNA of all genotypes except of the PTC1 homozygote. The HBA-T1 cDNA served as a RT–PCR control and was amplified in all individuals. (c) Urea cellulose acetate electrophoresis of globin polypeptides demonstrating the absence of a HBA-T3 translation product in the homozygotes for both PTC-containing alleles, whereas the heterozygotes and the individuals with non-PTC genotypes show a detectable (although weakly staining) band (arrows).