Figure 5
Transcripts detection in wt and splicing mutation minigenes. A) Fluorescent capillary electrophoresis of RT-PCR products generated by the wild type, c.-5 + 2dupT and c.-5 + 1G > A minigenes. Screenshots of Peak Scanner electropherograms are shown. Fluorescent RT-PCRs (blue peaks) of wt and mutant minigenes were run in an ABI3130 DNA sequencer with Genescan ROX 500 (red peaks) as size standard. RFU: Relative Fluorescence Units. B) Quantification of all detected of transcripts (Tr1 to 12) generated by the wild type, or mutants c.-5 + 1G > A (QOPorto) and c.-5 + 2dupT (QOMadrid) minigenes of the SERPINA1 gene are represented with the mean proportion of each one. Sizes were calculated by the Peak Scanner software. Depending the use of the alternative splicing sites described for exon 1B and 1C, the deduced transcript composition is: Tr1: V1-1Cs -V2; Tr2: V1-1Cl -V2; Tr3: V1-1Bs -V2; Tr4: V1-1Bl -V2; Tr5: V1-1Bs -1Cs-V2; Tr6: V1-1Bs -1Cl-V2; Tr7: V1-1Bl -1Cs-V2; Tr8: V1-1Bl -1Cl-V2; Tr9-12: partial intron retentions. (1Bs and 1Bl: exon1B short and long, respectively; 1Cs and 1Cl: exon 1C short and long, respectively).