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Single‐strand conformation polymorphism (SSCP) analysis of the molecular pathology of hemophilia B
Author(s) -
David Dezso,
Rosa Humberto A. V.,
Pemberton Susan,
Diniz Maria J.,
Campos Manuel,
Lavinha João
Publication year - 1993
Publication title -
human mutation
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.981
H-Index - 162
eISSN - 1098-1004
pISSN - 1059-7794
DOI - 10.1002/humu.1380020506
Subject(s) - single strand conformation polymorphism , biology , genetics , microbiology and biotechnology , intron , gene duplication , polymerase chain reaction , haemophilia b , gene , polymorphism (computer science) , dna , mutation , dna sequencing , sequence analysis , genotype , haemophilia a , haemophilia
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 FIX Barcelos Thr‐380‐Pro and FIX Lousada 9bp insertion at position 31,309 or 31,318; five mutations previously reported in other ethnic groups (FIX Porto Arg‐145‐His, FIX Luanda Gly‐207‐Arg, FIX Penafiel Arg‐248‐Gln, FIX Sesimbra Arg‐333‐Gln, FIX Cascais 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 (FIX Lousada ). We have performed molecular modeling studies in order to predict the structure of the variant FIX molecules. © 1993 Wiley‐Liss, Inc.