N1303K (c.3909C>G) Mutation and Splicing: Implication of Its c.[744-33GATT(6); 869+11C>T] Complex Allele in CFTR Exon 7 Aberrant Splicing

Figure 3

Impact of the c.[744-33GATT(6); 869+11C>T] complex allele on splicing patterns. (a) Polyacrylamide gel showing, for each plasmid construction, the different fragments of cDNA using 2,3α and Bra2rev primers on the total cDNA. This polyacrylamide gel shows three independent transfections of the c.[744-33GATT(7); 869+11C] (Lines 1, 2, and 3) and c.[744-33GATT(6); 869+11C>T] (Lines 5, 6, and 7). The results were confirmed in three different cell lines. Two fragments are visible: normal splicing with exon 7 (375 pb) and exon 7 skipping (249 pb). Ladder: ϕX174 DNA/BsuRI (HaeIII) Marker (Fermentas) was used (Lines 4 and 8). (b) Sequencing of the 375 bp fragment revealing a normal splicing and that of the 249 bp fragment revealing an exon 7 skipping.

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