Abstract
Ten to fifteen percent of CF chromosomes carry mutations which are not detected by routine screening of the CFTR gene for known mutations. Many techniques have been used to screen the CFTR gene for these remaining mutations. Most of the methods use genomlc DNA, and since the CFTR gene contains 27 exons, are necessarily labour intensive. We have screened the entire coding region of CFTR, by chemical cleavage of 7 overlapping segments of amplified cDNA. Using this method we have identified 4 sequence changes which had not been detected by screening genomlc DNA, and successfully detected 10 out of 13 known mutations. In addition, we have Identified 8 alternatively spliced forms of CFTR mRNA, 4 of which have not been described previously. These include transcripts lacking a) exon 3, b) exons 2 + 3, c) exons 9 + 12, and d) the final 357 bp of exon 15 as a result of use of the cryptic splice donor site CA2863/ GTTCGT).
| Original language | English (US) |
|---|---|
| Pages (from-to) | 1141-1146 |
| Number of pages | 6 |
| Journal | Human molecular genetics |
| Volume | 3 |
| Issue number | 7 |
| DOIs | |
| State | Published - Jul 1994 |
Funding
The authors are grateful to the CF patients who provided samples for this study, to Dr A.Thomson, Dr M.Super arid Dr J.Stroobant, and for allowing access to their patients and to Dr A.Sellar at the Churchill Hospital, Oxford, S.Dear at UMDS Guy's Hospital, London and Dr M.Schwartz at Royal Manchester Children's Hospital, for preliminary genotyping. The work was supported by the Cystic Fibrosis Trust and the Medical Research Council. J.H. is also supported by the H.C.Roscoe Fellowship from the BMA.
ASJC Scopus subject areas
- Genetics(clinical)
- Genetics
- Molecular Biology