ABCMdb

PMID: 1384328

Abeliovich D, Lavon IP, Lerer I, Cohen T, Springer C, Avital A, Cutting GR
Screening for five mutations detects 97% of cystic fibrosis (CF) chromosomes and predicts a carrier frequency of 1:29 in the Jewish Ashkenazi population.
Am J Hum Genet. 1992 Nov;51(5):951-6., [PubMed]

Sentences

No. Mutations Sentence Comment

33 ABCC7 p.Trp1282* ABCC7 p.Asn1303Lys ABCC7 p.Gly542* The regions encompassing the mutations of interest were simultaneously amplified by PCR using DNA primers: C16B and C16D for AF508 (Kerem et al. 1989), lli5 and 11i3 for G542X, 20i5 and 20i3 for W1282X, and 21i5 and 21i3 for N1303K (Kerem et al. 1990; Vidaud et al. 1990; Osborne et al. 1991; Zielenski et al. 1991 b). Login to comment 37 ABCC7 p.Gly542* Mutation Detection Mutations AF508 and G542X were detected by using oligonucleotides oligo-N, oligo AF, 542NL, and 542X according to a method described elsewhere (Kerem et al. 1989 and 1990). Login to comment 38 ABCC7 p.Trp1282* Mutation W1282X was detected by oligonucleotide 1282X (5'-CAACA- GTGAAGGAAAGCCTT-3'), while the oligonucleotide 1282NL (5'-CAACAGTGGAGGAAAGCCTT- 3'), was used to detect the normal allele. Login to comment 42 ABCC7 p.Trp1282* ABCC7 p.Gly542* The hybridization and wash conditions were as follows: W1282X-hybridization at 370C for 60 min and three washes in 2 x SSC, 0.1% SDS (twice at room temperature for 10 min and then at 590C for 10 min); AF508-hybridization 370C for 60 min and then three washes in 2 x SSC, 0.1% SDS (twice at 370C for 10 min and then at 420C for 10 min); G542X-hybridization at 420C for 2 h and then two washes in 2 x SSC, 0.1% SDS (once at 420C for 10 min and once at 450C for 10 min). Login to comment 43 ABCC7 p.Trp1282* The W1282X mutation was also identified by cleavage of the PCR product with MnlI (Vidaud et al. 1990). Login to comment 44 ABCC7 p.Asn1303Lys The mutation N1303K was detected by PCR-mediated site-directed mutagenesis as described by Friedman et al. Login to comment 46 ABCC7 p.Gly542* The same approach was used as an alternative method to detect AF508 and G542X. Login to comment 50 ABCC7 p.Gly551Asp ABCC7 p.Ser549Arg ABCC7 p.Ser549Asn The mutations G551D, S549N, S5491, S549R, and 1717-1 G-OA were analyzed according to the method of Kerem et al. Login to comment 51 ABCC7 p.Arg553* (1990), R553X according to the method of Cutting et al. Login to comment 54 ABCC7 p.Trp1282* ABCC7 p.Asn1303Lys ABCC7 p.Gly542* Results Five mutations-W1282X, AF508, G542X, N1303K, and 3849 + lOkb C-'lT-were found in our patients. Login to comment 56 ABCC7 p.Gly551Asp ABCC7 p.Ser549Arg ABCC7 p.Ser549Asn ABCC7 p.Ser549Ile The mutations 621 + 1 G-oT, 1717-1 G-oA, S549N, S549I, S549R, G551D, and R553Xwere not foundin our patients. Login to comment 57 ABCC7 p.Trp1282* ABCC7 p.Asn1303Lys ABCC7 p.Gly542* Amongthe Ashkenazi, three mutations-W1282X (48%), AF508 (30%), and G542X (12%)-accounted for 90% of the CF mutations, while mutations N1303K and 3849 + 1Okb C--T were found on an additional 7% of CF chromosomes. Login to comment 58 ABCC7 p.Trp1282* In the non-Ashkenazi Jews, the AF508 mutation accounted for 43% of the CF mutations; however, the W1282X (3%) mutation was identified on only one CF chromosome, leaving 54% of the CF mutations unidentified. Login to comment 60 ABCC7 p.Trp1282* ABCC7 p.Asn1303Lys ABCC7 p.Gly542* In the Arab patients, four mutations-AF508, N1303K, W1282X, and G542X-were found, accounting for 55% ofthe CF mutations. Login to comment 61 ABCC7 p.Asn1303Lys The N1303K mutation was more frequent in the Arabs than in the Jewish patients. Login to comment 62 ABCC7 p.Trp1282* ABCC7 p.Trp1282* ABCC7 p.Gly542* To evaluate the gene frequency in the Ashkenazi population, a random sample of 424 Ashkenazi individuals (848 chromosomes) was screened for the presence of the three mutations W1282X, AF508, and G542X. Thirteen individuals were identified as heterozygotes- six with the AF508 mutation and seven with the W1282X mutation. Login to comment 63 ABCC7 p.Gly542* In this sample we did not identify any carrier having the G542X mutation. Login to comment 64 ABCC7 p.Trp1282* ABCC7 p.Asn1303Lys ABCC7 p.Gly542* Discussion The predominant mutation in Caucasians, AF508, was found in all ethnic communities in Israel but at a significantly lower frequency (31%) than in northern Table 2 Distribution of CF Mutations, by Ethnic Group FREQUENCY (no.) IN Jews Ashkenazi Non-Ashkenazi Turkish Arabs MUTATION (n = 94) (n = 37) (n = 7) (n = 31) F508 ........................... .30 (28) .43 (16) .29 (2) .26 (8) G542X .................. .12 (11) .43 (3) .03 (1) W1282X ................... .48 (45) .03 (1) .14 (1) .10 (3) N1303K .................. .03 (3) .16 (5) 3849 + 10kb C-T .......... .04 (4) .14 (1) Subtotal .................. .97 .46 1.00 .55 Unidentified mutations .... .03 (3) .54 (20) 0 .45 (14) NOTE. Login to comment 82 ABCC7 p.Trp1282* ABCC7 p.Gly542* We screened for three mutations-AF508, W1282X, and G542X-which are expected to detect 90% of the carriers. Login to comment 84 ABCC7 p.Trp1282* In this sample, 13 individuals were detected as heterozygotes for either the W1282X mutation or the AF508 mutation. Login to comment