Table 3.
Effect of DNAH11 splice mutations on cDNA transcript using Reverse Transcriptase PCR (RT-PCR) in patients with PCD
| Sample # | Intron/Exon Location | Genomic Mutations and Predicted Amino-Acid Change | cDNA Transcript after RT-PCR | Comments |
|---|---|---|---|---|
| OP41-II:1 | Exon I | c.350A>T (p.E117V) Splice defect? |
r.(spl?) RNA not available |
Second last base in exon 1 on conserved canonical splice donor site. Population studies: 0/216 control alleles and 1/326 PCD alleles |
| PCD761 | Intron 13 | c.IVS13-1G>C (c.2275-1G>C) Splice defect |
r.2275_2667del p.Y759_JE889del |
Inframe deletion of exon 14 consisting of 131 amino-acid residues Wild type amplification product: 1089 bp Mutant amplification product: 696 bp |
| OP406-II:2 | Intron 23* | c.IVS23+5G>T (c.4254+5G>T) Splice defect |
r.4096_4254del p.E1366_G1418del |
Inframe deletion of exon 23 consisting of 53amino-acid residues Wild type amplification product: 741 bp Mutant amplification product: 582 bp |
| OP406-II:2 | Intron 26 | c.IVS26-1G>A (c.4726-1G>A) Splice defect |
r.4726_4817del p.E1576AfsX4 |
Out-of-frame deletion of exon 27 leading to premature translation termination signal Wild type amplification product: 992 bp Mutant amplification product: 900 bp |
| PCD653† | Intron 33* | c.IVS33+1G>A (c.5778+1G>A) Splice defect |
r.5461_6041del p.V1821TfsX7 |
Out-of-frame deletion of exons 32–35 leading to premature translation termination signal Wild type amplification product: 1013 bp Mutant amplification product: 432 bp |
| PCD108 | Intron 44 | c.IVS44+1G>A (c.7266+1G>A) Splice defect |
r.7135_7266del p.T2379_Q2422del |
Inframe deletion of exon 44 consisting of 44 amino-acid residues Wild type amplification product: 918 bp Mutant amplification product: 786 bp |
| OP98-II:1 | Exon 48 | c.7914G>C (p.Q2638H) Splice defect |
r.7812_7914del p.W2604X |
Last base in exon 48 on conserved canonical splice donor site. Out-of-frame deletion of exon 48 leading to premature translation termination signal Wild type amplification product: 1090 bp Mutant amplification product: 987 bp |
*
Intron 23 and Intron 33 analysis showed the absence of last 15 bases (5 amino-acid residues) in exon 22 and 6 bases of exon 32 (2 amino-acid residues) respectively, in multiple controls depicting error in published sequence.
†
RNA from affected individual PCD565 was not available hence cDNA analysis was done on the carrier father (PCD653).