ABCMdb

PMID: 15727254

Hu X, Plomp A, Gorgels T, Brink JT, Loves W, Mannens M, de Jong PT, Bergen AA
Efficient molecular diagnostic strategy for ABCC6 in pseudoxanthoma elasticum.
Genet Test. 2004 Fall;8(3):292-300., [PubMed]

Sentences

No. Mutations Sentence Comment

4 ABCC6 p.Arg1141* The two most frequent mutations, R1141X and an ABCC6 del exons 23-29, as well as a core set of mutations, were identified by restriction enzyme digestion and size separation on agarose gels. Login to comment 57 ABCC6 p.Arg1141* ABCC6 p.Gly1302Arg ABCC6 p.Arg765Gln ABCC6 p.Ala1303Pro ABCC6 p.Arg1459Cys Six nucleotide changes, including 2,294g Ǟ a (R765Q in exon 18), 3,421c Ǟ t (R1141X in exon 24), 3904g Ǟ a (G1302R in exon 28), 3,907g Ǟ c (A1303P in exon 28), 3,775del T (W1259 frameshift in exon 27), and 4,377c Ǟ t (R1459C in exon 30) were determined by the digestion of PCR fragments with restriction enzymes Sma I, Bsi YI, Nci I, Hea III, Bst NI, and Aci I, respectively. Login to comment 95 ABCC6 p.Arg1141* ABCC6 p.Gly1302Arg ABCC6 p.Arg765Gln ABCC6 p.Ala1303Pro ABCC6 p.Thr1130Met ABCC6 p.Arg1459Cys ABCC6 p.Arg1221Cys ABCC6 p.Gln749* ABCC6 p.Arg1114His ABCC6 p.Lys1394Asn ABCC6 p.Asp1326Asn RESULTS OF THE MUTATION ANALYSIS IN THE ABBC6 GENE IN 76 PATIENTS Sequence Type AA change variation Location Alleles Statusa Phaseb Method Nonsense Q749X 2247 C → T Exon 17 2 ht 3 DHPLC R1141X 3421 C → T Exon 24 35 hm.ht.ch 1 Bsi YI Missense R765Q 2294G → A Exon 18 1 ht 2 Sma I R1114H 3341G → A Exon 24 1 ht 3 dHPLC T1130M 3390C → T Exon 24 2 ch 3 dHPLC R1221C 3663C → T Exon 26 1 ch 3 dHPLC G1302R 3904G → A Exon 28 1 ht 2 Nci I A1303P 3907G → C Exon 28 1 ch 2 Hae III D1326N 3999G → A Exon 28 1 ht 3 dHPLC K1394N 4182G → T Exon 29 1 ch 3 dHPLC R1459C 4377C → T Exon 30 1 ht 2 Aci I Frameshift Splicing IVS17-12delTT Intron 17 1 ht 3 dHPLC IVS26-1G → A Intron 26 1 ht 3 dHPLC Deletion 1944del22 Exon 16 2 ht,ch 2 PCR 4182delG Exon 29 6 hm,ht 3 dHPLC 3775delT Exon 27 11 hm,ht 2 Bst NI 3821del48 Exon 27 1 ht 2 PCR Insertion 4220insAGAA Exon 30 1 ht 3 dHPLC Deletion A995del405 del exon 23-29 Exon 23-29 17 hm,ht,ch,hem 1 PCR ABCC6 2 ht 4 FISH ahm, homozygote; ht, heterozygote; ch, compound heterozygote; hem, hemizygote. Login to comment 98 ABCC6 p.Arg1141* ABCC6 mutation analysis. Phase I: Detection of the R1141X mutation in exon 24 and deletion of exons 23-29. Login to comment 100 ABCC6 p.Arg1141* Lane 1, the R1141X mutation disrupts the Bsi YI site in the wild-type sequence and results in a single PCR fragment of 100 bp; lane 2, the wild-type allele was cleaved into two Bsi YI fragments of 88 bp and 12 bp (latter band not visible). Login to comment 115 ABCC6 p.Arg1141* Phase 1: Rapid detection of the founder mutation R114X and deletion of exons 23-29 The founder mutation, R1141X (Hu et al., 2003b), was found in 35 out of 89 mutant alleles (39.3%). Login to comment 116 ABCC6 p.Arg1141* R1141X was the most common PXE mutation in our cohort. Login to comment 154 ABCC6 p.Arg1141* Phase 2: Rapid detection of a core set of mutations After screening of R1141X and del exons 23-29, a core set of seven additional mutations was rapidly identified by PCR and PCR-RFLP. Login to comment 157 ABCC6 p.Gly1302Arg ABCC6 p.Arg765Gln ABCC6 p.Ala1303Pro ABCC6 p.Arg1459Cys Next, R765Q, G1302R, A1303P, 3775 del T, and R1459C mutations were detected in 15 mutant alleles by digestion of PCR fragments with restriction enzymes Sma I, Nci I, Hae III, Bst NI, and Aci I, respectively. Login to comment 162 ABCC6 p.Thr1130Met ABCC6 p.Arg1221Cys ABCC6 p.Gln749* ABCC6 p.Arg1114His ABCC6 p.Lys1394Asn ABCC6 p.Asp1326Asn Ten additional known nucleotide changes, including Q749X, R1114H, T1130M, R1221C, D1326N, K1394N, 4182delG, IVS17-12delTT, IVS26-1G Ǟ A, and 4220ins AGAA were successfully identified by dHPLC under the conditions presented in Table 1 (Fig. 3). Login to comment 172 ABCC6 p.Arg1141* The current strategy is especially efficient for the European population, in which a number of PXE mutations occur very frequent, i.e., R1141X (39.3%) and del exon 33-29 (19%). Login to comment 196 ABCC6 p.Asp1326Asn In addition, two additional novel mutant alleles (IVS 26-1 g Ǟ a, D1326N), which were missed by SSCP analysis previously, were identified. Login to comment 214 ABCC6 p.Arg1459Cys Our group investigated one family with a missense mutation (R1459C) by genotype, transcript, or haplotype and found no mutation on the second allele of the ABCC6 gene (Hu et al., 2003a; Plomp et al., 2004). Login to comment