ABCMdb

PMID: 19710401

El-Seedy A, Dudognon T, Bilan F, Pasquet MC, Reboul MP, Iron A, Kitzis A, Ladeveze V
Influence of the duplication of CFTR exon 9 and its flanking sequences on diagnosis of cystic fibrosis mutations.
J Mol Diagn. 2009 Sep;11(5):488-93., [PubMed]

Sentences

No. Mutations Sentence Comment

32 ABCC7 p.Asp443Tyr Surprisingly, only one mutation was identified in her father (p.Asp443Tyr in exon 9), and no mutation was found in her mother. Login to comment 33 ABCC7 p.Asp443Tyr The same result (p.Asp443Tyr mutation) was obtained in Bordeaux with case 1. Login to comment 36 ABCC7 p.Asp443Tyr The new analysis for this family confirmed the previous results: case 1 carries the c.1392 ϩ 6insC and c.1392 ϩ 12GϾA mutations in intron 9, but not the Asp443Tyr mutation in exon 9. Login to comment 37 ABCC7 p.Asp443Tyr On analysis of fresh parental samples at Bordeaux Hospital, the previous parental results were also confirmed, showing that her father must be carrier for Asp443Tyr mutation in exon 9. Login to comment 67 ABCC7 p.Asp443Tyr Sequencing of the PCR fragments using CF9.6 and CF9 ϩ 58 primers revealed two heterozygous mutations, c.1392 ϩ 6insC and c.1392 ϩ 12GϾA. However, PCR using the CF9-130 and CF9 ϩ 58 primers (PCR product B, Bordeaux primers), revealed only the GϾT variation at position 1327, corresponding to the p.Asp443Tyr mutation, and not the expected c.1392 ϩ 6insC and c.1392 ϩ 12GϾA mutations. Login to comment 70 ABCC7 p.Asp443Tyr Surprisingly, only the p.Asp443Tyr mutation was detected using another primers pair that spans from intron 8 to intron 9, (CF9-268 and CF9 ϩ 121, PCR C, from intron 8 to intron 9 as for Bordeaux primers). Login to comment 72 ABCC7 p.Asp443Tyr The results revealed that PCR product F covers only the Asp443Tyr mutation, whereas PCR products obtained using exonic Table 1. Login to comment 85 ABCC7 p.Asp443Tyr Sequencing analysis of cloned PCR product C showed two alleles: one mutated (p.Asp443Tyr allele) and one wild-type. Login to comment 99 ABCC7 p.Asn416Ser ABCC7 p.Asn416Ser ABCC7 p.Lys464Asn ABCC7 p.Lys464Asn Several variations have been identified in exon 9, including four possible mutations shown in Figure 2A: p.Lys464Asn, c.1328_1329delAT, c.1235delC, and p.Asn416ser (http://www.genet.sickkids.on.ca/cftr/; last accessed 24/12/2008) [Personal communications: p.Lys464Asn (E. Bleth, V. Gaston, P. Gautry), c.1328_1329delAT (T. Bienvenu, L. Tchertkoff, C. Cazeneuve, C Beldjord), c.1235delC (C. Fe´rec), and p.Asn416Ser (L. Picci, M. Cameran, O. Marangon, D. Marzenta, M. Scarpa)]. Login to comment 105 ABCC7 p.Asp443Tyr ABCC7 p.Asp443Tyr Summary of Clones with Different Genotypes Used for PCR Amplification and Sequencing Case Clone genotype N/N N/N c.1327GϾT(p.Asp443Tyr) N/N N/N c.1392 ϩ 6insC; c.1392 ϩ 12GϾ A c.1327GϾT(p.Asp443Tyr) c.1392 ϩ 6insC; c.1392 ϩ 12GϾ A Total number of clones tested Case 1 % clones of PCR A 19 54 27 0 26 % clones of PCR C 60 40 0 0 15 Case 2 % clones of PCR A 4 26 35 35 15 % clones of PCR C 67 33 0 0 6 N/N ϭ wild-type genotypes. Login to comment 114 ABCC7 p.Asp443Tyr Moreover, when the annealing temperature of PCR A was increased from 50°C to 60°C, the PCR product A presented only the p.Asp443Tyr mutation (GϾT at 1327) in exon 9, and not the c.1392 ϩ 6insC; c.1392 ϩ 12GϾA mutations. Login to comment 133 ABCC7 p.Lys464Asn ABCC7 p.Lys464Asn ABCC7 p.Lys464Asn Mutations in CFTR Exon 9 and its Intronic Boundaries that Have Homologous Sequences in Other Chromosomes CFTR mutation Common nomenclature Nucleotide change Site of mutation* Consequences p.Lys464Asn K464N G to T at 1392 Exon 9 (no. 10) Lys to Asn at 464; mRNA splicing defect? Login to comment 134 ABCC7 p.Asn416Ser ABCC7 p.Asn416Ser ABCC7 p.Asn416Ser c.1328_1329delAT 1460delAT Deletion of AT from 1328 Exon 9 (no. 10) Frameshift c.1235delC 1367delC Deletion of C at 1235 Exon 9 (no. 10) Frameshift p.Asn416Ser N416S A to G at 1247 Exon 9 (no. 10) Asn to Ser at 416 c.1392 ϩ 6insC; c.1392 ϩ 12GϾA 1524 ϩ 6insC 1524 ϩ 12GϾA Insertion of C after 1392 ϩ 6, G to A at 1392 ϩ 12 Intron 9 (no. 10) mRNA splicing defect? Login to comment 145 ABCC7 p.Asn416Ser ABCC7 p.Lys464Asn These include p.Lys464Asn, c.1328_1329delAT, c.1235delC, and p.Asn416Ser in exon 9, and c.1392 ϩ 6insC; c.1392 ϩ 12GϾA in intron 9. Login to comment