Tablel 3

Effect of DNAH11 splice mutations on cDNA transcript using reverse transcriptase PCR (RT-PCR) in patients with primary ciliary dyskinesia (PCD)

Sample no.	Intron/exon location	Genomic mutations and predicted amino acid change	cDNA transcript after RT-PCR	Comments
OP41-II:1	Exon 1	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_E889del)	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 53 amino 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 (five amino acid residues) in exon 22 and six bases of exon 32 (two 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).
DNAH11, dynein axonemal heavy chain 11.