ABCMdb

PMID: 1370807

Plieth J, Rininsland F, Schlosser M, Cooper DN, Reiss J
Single-strand conformation polymorphism (SSCP) analysis of exon 11 of the CFTR gene reliably detects more than one third of non-delta F508 mutations in German cystic fibrosis patients.
Hum Genet. 1992 Jan;88(3):283-7., [PubMed]

Sentences

No. Mutations Sentence Comment

16 ABCC7 p.Gly551Asp ABCC7 p.Arg553* ABCC7 p.Gly542* Indeed, two mutational "hot spots" have been identified, one of which, located within the 95-bp exon 11 (Cutting et al. 1990), contains a number of mutations including G551D, R553X, 1717-1G--~A (splice-site mutation), G542X and $549R. Login to comment 18 ABCC7 p.Gly551Asp ABCC7 p.Arg553* ABCC7 p.Gly542* A combination of restriction enzyme digestion (for R553X and G551D) and/or allele-specific oligonucleotide hybridization (for G542X and 1717-1G--~A) is now routinely performed to detect known mutations in polymerase chain reaction (PCR) amplified DNA samples. Login to comment 47 ABCC7 p.Gly551Asp ABCC7 p.Arg553* ABCC7 p.Gly542* Nearly half of these chromosomes contained R553X but the remainder contained G551D, G542X, 1717-1G--~A and $549R. Login to comment 54 ABCC7 p.Gly551Asp ABCC7 p.Arg553* ABCC7 p.Gly542* Distribution of non-AF508mutations in 59 German patients Non-AF508mutations Chromo- % (n = 72) somes In exon 11 R553X 12 16.6 G542X 5 6.9 G551D 4 5.5 1717-1G---~A(5' splicesite) 3 4.1 $549R 2 2.8 Others 9 12.5 Undetermined 37 51.4 Fig. la-c. Login to comment 59 ABCC7 p.Gly551Asp ABCC7 p.Arg553* ABCC7 p.Arg553* ABCC7 p.Arg553* ABCC7 p.Gly542* ABCC7 p.Gly542* W Wildtype control, lane 1 $549R/ $549R, lane 2 G551D, lane 3 1717-1G---~A,lane 4 G542X, lane 5 R553X, lane 6 R553X/R553X, lane 7 G542X/1717-1 SSCP analysis was then performed upon the same set of DNA samples to test the mutation-detection efficiency of this method. Login to comment 63 ABCC7 p.Gly551Asp ABCC7 p.Arg553* ABCC7 p.Gly542* Mutation-specific bands can thus be noted in patient samples bearing the heterozygous mutations G551D, 1717-1G--+A, G542X and R553X. Login to comment 64 ABCC7 p.Arg553* ABCC7 p.Gly542* Furthermore, individuals homozygous for the mutations $549R and R553X, and compound heterozygous for G542X/1717-1G-->A were readily distinguishable from patients heterozygous for these mutations because of the absence of wildtype bands in the former (Fig. 1). Login to comment 72 ABCC7 p.Gly551Asp ABCC7 p.Arg553* ABCC7 p.Gly542* ABCC7 p.Ser549Arg SSCP analysis performed on DNAs of six individuals with the following genotypes (nucleotide change in brackets): lanes W wildtype/wildtype, lanes 1 G551D/wildtype (G-*A), lanes 2 1717- 1G-~A/wildtype (G--~A), lanes 3 G542X/wildtype (G--~T), lanes 4 R553X/wildtype (C----~T),lanes 5 S549R/S549R(T-*G). Login to comment 75 ABCC7 p.Gly551Asp ABCC7 p.Gly542* In the case of the mutations G551D, G542X and $549, the coding strands of mutant and wild-type allele are separated, whereas the non-coding strands co-migrate. Login to comment 76 ABCC7 p.Arg553* In the case of the mutations 1717-1G--*A and R553X, the coding strands exhibit the same electropboretic mobility and only the non-coding strands of the wild-type and mutant allele can be distinguished lead to similar electrophoretic mobility of the mutation-bearing and wildtype strands. Login to comment