title: Identification of mutations leading to the Lesch-Nyhan syndrome by automated direct DNA sequencing of in vitro amplified cDNA.
abstract: The Lesch-Nyhan (LN) syndrome is a severe X chromosome-linked disease that results from a deficiency of the purine salvage enzyme hypoxanthine phosphoribosyltransferase (HPRT). The mutations leading to the disease are heterogeneous and frequently arise as de novo events. We have identified nucleotide alterations in 15 independently arising HPRT-deficiency cases by direct DNA sequencing of in vitro amplified HPRT cDNA. We also demonstrate that the direct DNA sequence analysis can be automated, further simplifying the detection of new mutations at this locus. The mutations include DNA base substitutions, small DNA deletions, a single DNA base insertion, and errors in RNA splicing. The application of these procedures allows DNA diagnosis and carrier identification by the direct detection of the mutant alleles within individual families affected by LN.
authors.affiliation: Baylor College of Medicine, Houston, TX.
authors.forename: R A
authors.initials: RA
authors.lastname: Gibbs
authors.forename: P N
authors.initials: PN
authors.lastname: Nguyen
authors.forename: L J
authors.initials: LJ
authors.lastname: McBride
authors.forename: S M
authors.initials: SM
authors.lastname: Koepf
authors.forename: C T
authors.initials: CT
authors.lastname: Caskey
mesh_terms: D000483:Alleles; D001322:Autoanalysis; D001483:Base Sequence; D004247:DNA / Q000235:genetics*; D004259:DNA-Directed DNA Polymerase; D005784:Gene Amplification; D006801:Humans; D007041:Hypoxanthine Phosphoribosyltransferase / Q000235:genetics*; D007926:Lesch-Nyhan Syndrome / Q000235:genetics*; D009154:Mutation; D009693:Nucleic Acid Hybridization; D015345:Oligonucleotide Probes; D010375:Pedigree; D012326:RNA Splicing; D012984:Software
chemical_list: D015345:Oligonucleotide Probes; D004247:DNA; D007041:Hypoxanthine Phosphoribosyltransferase; D004259:DNA-Directed DNA Polymerase