title: Mutation analysis of the hamartin gene using denaturing high performance liquid chromatography.
abstract: Denaturing high performance liquid chromatography (DHPLC) is a novel high-capacity technique for gene mutation scanning. We have assessed the sensitivity and specificity of this method for analysis of the full coding sequence of the hamartin (TSC1) gene in 20 tuberous sclerosis patients, whose TSC1 genes previously had been studied by single strand conformation polymorphism analysis and protein truncation assay. All eight sequence variants previously identified were adequately detected by DHPLC. Additionally, this approach picked up three polymorphisms, one of which (IVS13-55 C>G) was hitherto unreported, therefore serving as proof of principle for this technique. Thus, DHPLC appears to be a highly sensitive method with advantages in terms of flexibility, fragments size analysis, cost and time and labor sparing, compared to classical approaches of mutation scanning.
authors.affiliation: Unité de Recherches INSERM U-393, Département de Génétique, H opital des Enfants-Malades, Paris, France.
authors.forename: P
authors.initials: P
authors.lastname: Bénit
authors.forename: A
authors.initials: A
authors.lastname: Kara-Mostefa
authors.forename: M
authors.initials: M
authors.lastname: Berthelon
authors.forename: K
authors.initials: K
authors.lastname: Sengmany
authors.forename: A
authors.initials: A
authors.lastname: Munnich
authors.forename: J P
authors.initials: JP
authors.lastname: Bonnefont
mesh_terms: D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004247:DNA / Q000097:blood; D004252:DNA Mutational Analysis; D005260:Female; D016147:Genes, Tumor Suppressor / Q000235:genetics*; D006801:Humans; D008297:Male; D009691:Nucleic Acid Denaturation / Q000235:genetics; D011110:Polymorphism, Genetic / Q000235:genetics; D011506:Proteins / Q000235:genetics*; D014402:Tuberous Sclerosis / Q000235:genetics*; D000077004:Tuberous Sclerosis Complex 1 Protein; D025521:Tumor Suppressor Proteins
chemical_list: D011506:Proteins; C000624650:TSC1 protein, human; D000077004:Tuberous Sclerosis Complex 1 Protein; D025521:Tumor Suppressor Proteins; D004247:DNA