title: Single-strand conformation polymorphism (SSCP) analysis of the molecular pathology of hemophilia B.
abstract: In the present study, we report the application of polymerase chain reaction-single-strand conformation polymorphism (PCR-SSCP) analysis to the screening of seven functionally important factor IX gene (FIX) regions (total length 2.66 kb) in 9 unrelated haemophilia B patients of Portuguese or African origin. In eight of the patients an altered migration pattern of single-stranded DNA was observed. Direct sequencing of the relevant DNA fragments unveiled the following sequence alterations: two novel mutations, namely FIXBarcelos Thr-380-Pro and FIXLousada 9bp insertion at position 31,309 or 31,318; five mutations previously reported in other ethnic groups (FIXPorto Arg-145-His, FIXLuanda Gly-207-Arg, FIXPenafiel Arg-248-Gln, FIXSesimbra Arg-333-Gln, FIXCascais Arg-333-Stop); and a normal variant, G-->T transvertion at position 6,596 in intron 2. We propose hypothetical models for the generation of the 9 bp duplication (FIXLousada). We have performed molecular modeling studies in order to predict the structure of the variant FIX molecules.
authors.affiliation: Laboratório de Genética Humana, Instituto Nacional de Saúde Dr Ricardo Jorge, Lisboa, Portugal.
authors.forename: D
authors.initials: D
authors.lastname: David
authors.forename: H A
authors.initials: HA
authors.lastname: Rosa
authors.forename: S
authors.initials: S
authors.lastname: Pemberton
authors.forename: M J
authors.initials: MJ
authors.lastname: Diniz
authors.forename: M
authors.initials: M
authors.lastname: Campos
authors.forename: J
authors.initials: J
authors.lastname: Lavinha
mesh_terms: D001483:Base Sequence; D004247:DNA / Q000235:genetics*; D004277:DNA, Single-Stranded / Q000235:genetics; D005164:Factor IX / Q000235:genetics*; D002836:Hemophilia B / Q000235:genetics*; D006801:Humans; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics; D009690:Nucleic Acid Conformation; D016133:Polymerase Chain Reaction / Q000379:methods; D011110:Polymorphism, Genetic / Q000235:genetics*
chemical_list: D004277:DNA, Single-Stranded; D005164:Factor IX; D004247:DNA