title: Three novel mutations in the cystic fibrosis gene detected by chemical cleavage: analysis of variant splicing and a nonsense mutation.
abstract: We have used the chemical cleavage mismatch technique to screen for mutations in the cystic fibrosis gene. Analysis of exons 10 and 11 in the first nucleotide binding fold led to the detection of several described mutations and two novel mutations, V520F and C524X. V520F results from a G-->T nucleotide substitution changing a valine to a phenylalanine residue, while C524X (a nonsense mutation), results from a C-->A transversion. A third novel mutation, Q1291H (G-->C), at the last nucleotide of exon 20, would substitute a histidine residue for glutamine. Further study, involving RNA based PCR, revealed that Q1291H was also a splice mutation. Both correctly and aberrantly spliced mRNAs are produced from the Q1291H allele. The incorrectly spliced product results from the use of a nearby cryptic splice site 29 bases into the adjacent intron.
authors.affiliation: Human Genetics Unit, University of Edinburgh, UK.
authors.forename: C T
authors.initials: CT
authors.lastname: Jones
authors.forename: I
authors.initials: I
authors.lastname: McIntosh
authors.forename: M
authors.initials: M
authors.lastname: Keston
authors.forename: A
authors.initials: A
authors.lastname: Ferguson
authors.forename: D J
authors.initials: DJ
authors.lastname: Brock
mesh_terms: D000328:Adult; D000595:Amino Acid Sequence; D001483:Base Sequence; D018118:Chloride Channels; D003550:Cystic Fibrosis / Q000235:genetics*; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D004848:Epithelium / Q000502:physiology; D005091:Exons; D006801:Humans; D007438:Introns; D007473:Ion Channels / Q000235:genetics; D008297:Male; D008565:Membrane Proteins / Q000235:genetics*; D008969:Molecular Sequence Data; D009154:Mutation; D009838:Oligodeoxyribonucleotides; D017354:Point Mutation; D016133:Polymerase Chain Reaction / Q000379:methods; D012313:RNA / Q000235:genetics / Q000302:isolation & purification; D012326:RNA Splicing
chemical_list: C505032:CFTR protein, human; D018118:Chloride Channels; D007473:Ion Channels; D008565:Membrane Proteins; D009838:Oligodeoxyribonucleotides; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D012313:RNA