ABCMdb

PMID: 11933191

Ravnik-Glavac M, Atkinson A, Glavac D, Dean M
DHPLC screening of cystic fibrosis gene mutations.
Hum Mutat. 2002 Apr;19(4):374-83., [PubMed]

Sentences

No. Mutations Sentence Comment

42 ABCC7 p.Gly551Asp ABCC7 p.Arg117His ABCC7 p.Trp1282* ABCC7 p.Arg553* ABCC7 p.Arg334Trp ABCC7 p.Arg347Pro ABCC7 p.Trp1316* ABCC7 p.Arg347Cys ABCC7 p.Asn1303Lys ABCC7 p.Gly542* ABCC7 p.Arg1162* ABCC7 p.Asp1270Asn ABCC7 p.Arg74Trp ABCC7 p.Arg352Gln ABCC7 p.Pro99Leu ABCC7 p.Gln493* ABCC7 p.Gly85Glu ABCC7 p.Ser549Asn ABCC7 p.Arg560Thr ABCC7 p.Arg75Gln ABCC7 p.Arg1070Gln ABCC7 p.Ser1235Arg ABCC7 p.Ala120Thr ABCC7 p.Asn1303His ABCC7 p.Tyr1092* ABCC7 p.Ser1255* ABCC7 p.His1085Arg ABCC7 p.Arg1066Leu ABCC7 p.Ile1234Val ABCC7 p.Arg117Cys ABCC7 p.Leu1077Pro ABCC7 p.Arg1066His ABCC7 p.Ile336Lys ABCC7 p.Leu719* ABCC7 p.Arg560Lys ABCC7 p.Ser492Phe ABCC7 p.Ala349Val ABCC7 p.Asp110His ABCC7 p.Gln1291Arg ABCC7 p.Ala559Thr ABCC7 p.Lys716* ABCC7 p.Gln151* ABCC7 p.His1054Asp ABCC7 p.Glu92* ABCC7 p.Trp57Gly ABCC7 p.Gly628Arg ABCC7 p.Gly1061Arg ABCC7 p.Gln685* The following mutations have been studied: exon 3: W57G, R74W, R75Q, G85E, 394delTT, 405+ 1G>A; exon 4: E92X, P99L, 441delA, 444delA, 457TAT>G, D110H, R117C, R117H, A120T, 541delC, 544delCA, Q151X, 621+1G>T, 662- 2A>C; exon 7: 1078delT, F331L, R334W, I336K, R347C, R347P, A349V, R352Q, 1221delCT; exon 10: S492F, Q493X, 1609delCA, deltaI507, deltaF508; exon 11: G542X, S549N, G551D, R553X, A559T, R560K, R560T; exon 13: K716X, Q685X, G628R, L719X; exon 17b: H1054D, G1061R, 3320ins5, R1066H, R1066L, R1070Q, 3359delCT, L1077P, H1085R, Y1092X; exon 19: R1162X, 3659delC, 3662delA, 3667del4, 3737delA, I1234V, S1235R, 3849G>A; exon 20: 3860ins31,S1255X,3898insC,3905insT,D1270N, W1282X, Q1291R; and exon 21: N1303H, N1303K, W1316X. Login to comment 69 ABCC7 p.Arg74Trp ABCC7 p.Trp57Gly profiles of two exon 3 mutations, W57G and R74W, did not resolve from those of the normal sample at the recommended temperature of 55°C (Fig. 2). Login to comment 81 ABCC7 p.Arg74Trp ABCC7 p.Trp57Gly Two mutations (W57G and R74W) were not distinguished from the control sample at the temperature recommended by the MELT program. Login to comment 100 ABCC7 p.Arg334Trp ABCC7 p.Arg347Cys ABCC7 p.Asp1270Asn ABCC7 p.Arg74Trp ABCC7 p.Gln493* ABCC7 p.Trp57Gly Optimization of Temperature (OTm) for Undetected Mutations Nucleotide RTm OTm Exon Mutation change (°C) (°C) 3 W57G 301 T>G 55 57 R74W 352 C>T 55 57 7 R334W 1132 C>T 58 60 R347C 1171 C>T 58 60 10 Q493X 609 C>T 55 56 20 3905 insT 3905 insT 55 56 D1270N 3940 G>A 57 58 RTm, recommended temperature by the MELT program; OTm, optimized temperature. Login to comment 129 ABCC7 p.Gly628Arg ABCC7 p.Gln685* Of those exon 13 mutations that were not detected at 60°C, G628R and Q685X (see Fig. 4) each lies within a domain with a melting temperature of 56°C. Login to comment 130 ABCC7 p.Gly628Arg ABCC7 p.Gln685* It is therefore understandable that G628R and Q685X did not resolve at 60°C. Login to comment 167 ABCC7 p.Arg334Trp In CFTR gene we detected one mutation, a C>T change at position 1132 (R334W), which was not resolved at RTm 56°C but was only detected at temperature 58°C and therefore can be used as one of the calibrator standard mutations. Login to comment 176 ABCC7 p.Arg334Trp Mutation R334W in exon 7 of theCFTR gene was detected at one temperature only and can thus serve as one of the mutation standards; 4) at present, it is not yet understood why the MELT algorithm prediction of melting temperature may appear underestimated, although the column oven calibration can be one of the reasons. Login to comment