ABCMdb

PMID: 7526925

Hull J, Shackleton S, Harris A
Analysis of mutations and alternative splicing patterns in the CFTR gene using mRNA derived from nasal epithelial cells.
Hum Mol Genet. 1994 Jul;3(7):1141-6., [PubMed]

Sentences

No. Mutations Sentence Comment

16 ABCC7 p.Gly178Arg We have detected 14 sequence changes in the CFTR gene using this technique, including 4 (G178R (8), 2184delA (Bozon, pes comm. Login to comment 31 ABCC7 p.Gly178Arg These sequence changes included 2 mutations, G178R (8) and 2184delA (Bozon, pes comm. Login to comment 33 ABCC7 p.Gly551Asp ABCC7 p.Trp1282* ABCC7 p.Gly542* ABCC7 p.Gly85Glu ABCC7 p.Gln220* ABCC7 p.Glu92Lys Of the 13 known sequence changes, 9 (G85E (8), E92K (10), Q220X (11), AF508 (1), G542X (12), G551D (13), 3659delC (12), W1282X (14), 4271delC (11)) were readily identified by •To whom correspondence should be addressed A-6 \ B c ~~i r D t 1 F 1 2 3 4 5 Ga 6b 7 8 9 1 0 1 1 1 2 13 14i 14bl 516 17a 17bl S 19 202122 23 24 MSD 1 NBF 1 R domain MSD 2 NBF 2 Figure 1. Login to comment 37 ABCC7 p.Gly551Asp ABCC7 p.Gly542* Autoradiograph showing cleavage products for the 3659delC mutation (577 bases) in the E fragment AF5O8 (634 + 267 bases), G542X (164 bases) and G551D (136 bases) mutations in the B fragment. Login to comment 43 ABCC7 p.Gly551Asp ABCC7 p.Gly542* An autoradiograph showing the labeled cleavage products for the AF508, G542X, G551D, and 3659delC mutations is seen in Figure 2. Login to comment 47 ABCC7 p.Arg553* ABCC7 p.Gly27* The 4 mutations that were not detected were 182delT (11), G27X (15), R553X (14) and W1O98R (Tsui pers comm). Login to comment 48 ABCC7 p.Trp1098Arg The first 3 of these mutations were readily detectable by chemical cleavage on amplified genomic DNA, but the fourth, W1098R, was not. Login to comment 51 ABCC7 p.Gly27* 182delT (11) G27X (15) G85E0 (8) £92*? Login to comment 53 ABCC7 p.Arg553* (1) G542XC (12) GSSID0 (13) R553X (13) W1098Rc (Tsui pers. Login to comment 58 ABCC7 p.Trp1098Arg for the W1098R mutation, in which the patient sample, rather than the control, was end labelled with [7-32 P] dATP and yielded a cleavage product of the predicted size (Figure 3). Login to comment 66 ABCC7 p.Trp1098Arg Of the 13 known sequence changes, wt labeled W1098R labeled 1 % i -342b Figure 3. Login to comment 67 ABCC7 p.Trp1098Arg Detection of the W1098R mutation. Login to comment 75 ABCC7 p.Arg553* ABCC7 p.Trp1098Arg ABCC7 p.Gly27* Mutations not detected by the chemical cleavage technique The mutations that this method failed to detect were 182delT (exon 1, A-6 set), G27X (G to T substitution, exon 2, A-6 set), R553X (C to T substitution, exon 11, B set), and W1098R (T to C substitution, exon 17b, E set). Login to comment 78 ABCC7 p.Arg553* The failure to detect the R553X mutation is also readily explained. Login to comment 82 ABCC7 p.Trp1098Arg ABCC7 p.Gly27* Analysis of the full length cDNA fragments A-6 and E respectively, by direct sequencing in the case of G27X, and by restriction enzyme digestion for the W1098R mutation (the T to C mutation creates a unique Mspl recognition site in the E fragment), demonstrated that both mutations were present in their respective full length fragments of amplified cDNA. Login to comment 85 ABCC7 p.Trp1098Arg The W1098R mutation (which was not detected by chemical mismatch analysis of exon 17b genomic DNA previously (11)), is caused by a T to C substitution and results in the formation of a T-G mismatch. Login to comment 100 ABCC7 p.Gly27* The G27X mutation results from a G to T change, producing a C-T mismatch which should be readily detectable by hydroxylamine. Login to comment 137 ABCC7 p.Gly551Asp ABCC7 p.Arg553* ABCC7 p.Gly542* They had been screened for the AF5O8, G542X, G551D, R553X and 621 + 1G-T mutations by the clinical genetics service at the Churchill Hospital, Oxford, and they did not carry these mutations. Login to comment