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Single-strand conformation polymorphism (SSCP) analysis of the molecular pathology of hemophilia B

1993; Wiley; Volume: 2; Issue: 5 Linguagem: Inglês

10.1002/humu.1380020506

1098-1004

D. David, Humberto A. V. Rosa, Susan Pemberton, Maria João Diniz, Manuel Campos, João Lavinha,

Biochemical and Molecular Research

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. © 1993 Wiley-Liss, Inc.