title: Screening for mutations in the phenylalanine hydroxylase gene from Italian patients with phenylketonuria by using the chemical cleavage method: a new splice mutation.
abstract: To investigate the molecular basis of phenylketonuria in Italy we applied the chemical cleavage method (CCM) on amplified DNA encompassing exons 7 and 8 of the phenylalanine hydroxylase gene. These exons are in a region likely to be involved in enzyme function. Using this approach, we could simultaneously screen for novel mutations and for seven reported mutations which map in this area. Three mutations were identified. The first was shown to be a not previously described mutation: a G----A substitution at the 5' donor junction splice site of intron 7. The second change was a reported G----A mutation at codon 261. The third change corresponded to a polymorphism at codon 245. Our results indicate that CCM analysis of amplified genomic DNA fragments can be successfully used to search for mutations in large genes whose transcripts are not readily available.
authors.affiliation: Istituto di Clinica Pediatrica, Centro di Immunogenetica e Istocompatibilita CNR, Turin, Italy.
authors.forename: I
authors.initials: I
authors.lastname: Dianzani
authors.forename: S M
authors.initials: SM
authors.lastname: Forrest
authors.forename: C
authors.initials: C
authors.lastname: Camaschella
authors.forename: G
authors.initials: G
authors.lastname: Saglio
authors.forename: A
authors.initials: A
authors.lastname: Ponzone
authors.forename: R G
authors.initials: RG
authors.lastname: Cotton
mesh_terms: D001483:Base Sequence; D004247:DNA / Q000096:biosynthesis; D005091:Exons; D005820:Genetic Testing; D006801:Humans; D007558:Italy; D008969:Molecular Sequence Data; D009154:Mutation; D010651:Phenylalanine Hydroxylase / Q000235:genetics*; D010661:Phenylketonurias / Q000175:diagnosis* / Q000235:genetics; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length
chemical_list: D004247:DNA; D010651:Phenylalanine Hydroxylase