Allelic Imbalance in TOR1A mRNA Expression in Manifesting and Non-Manifesting Carriers of the GAG-Deletion

Figure 1

Primers used in allele-specific PCR assays: partial sequence of the 4th and 5th exons of human torsinA is shown and their intronic sequence is indicated by “-INTRON-.” The primer pair (TOR1A-F and TOR1A-R) used to generate a 327 bp human TOR1A fragment from RNA is shown in bold italics. The allele-specific pairs of primers used to discriminate between the WT and ΔE TOR1A alleles are underlined. The same allele-specific forward primer is used (AS-F) with two different reverse primers, one matching the WT allele with 3′ ending at CTCCTC (ASWT-R) and the other matching the mutant allele, missing a 303 bp-GAG and thus with a 3′ ending at CGACTC (ASΔE-R). The last three nucleotides, which are different between the two reverse primers, are highlighted in grey; the dashes represent the 3 nucleotides deleted in the ASΔE-R primer. The stop codon (SC) TGA is also indicated in bold. All reverse primers are shown under their corresponding forward sequence in reverse complement orientation.