Application of natural and amplification created restriction sites for the diagnosis of PKU mutations.
PCR amplification, either conventional, or as site directed mutagenesis using primers with mismatched 3'-ends, followed by restriction endonuclease digestion, provides rapid, non-isotope assays of known mutations in the human phenylalanine hydroxylase gene. Such assays were shown to have the potential to detect all of the 18 presently reported phenylketonuria mutations. The practical applicability of this approach was demonstrated for eight mutations in Norwegian phenylketonuria patients, among them the most common ones.
Eiken
H G
HG
Department of Medical Genetics, Haukeland Hospital, University of Bergen, Norway.
Odland
E
E
Boman
H
H
Skjelkvåle
L
L
Engebretsen
L F
LF
Apold
J
J
D001483:Base Sequence; D004262:DNA Restriction Enzymes; D004587:Electrophoresis, Agar Gel; D005784:Gene Amplification; D006801:Humans; D008969:Molecular Sequence Data; D009154:Mutation; D010651:Phenylalanine Hydroxylase / Q000235:genetics*; D010661:Phenylketonurias / Q000175:diagnosis / Q000235:genetics*; D016133:Polymerase Chain Reaction; D015183:Restriction Mapping
D010651:Phenylalanine Hydroxylase; D004262:DNA Restriction Enzymes