Mutations and a polymorphism in the factor VIII gene discovered by denaturing gradient gel electrophoresis

Journal Article · Thu Mar 01 04:00:00 EST 1990 · Proceedings of the National Academy of Sciences of the United States of America; (USA)

DOI:https://doi.org/10.1073/pnas.87.6.2092· OSTI ID:6074714

Kogan, S; Gitschier, J ^[1]

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1. Univ. of California, San Francisco (USA)

Hemophilia A results from mutations in the gene coding for coagulation factor VIII. The authors gradient gel electrophoresis to screen for mutations in the region of the factor VIII gene coding for the first acidic domain. Amplification primers were designed employing the MELTMAP computer program to optimize the ability to detect mutations. Screening of amplified DNA from 228 unselected hemophilia A patients revealed two mutations and one polymorphism. Rescreening the same population by making heteroduplexes between amplified patient and control samples prior to electrophoresis revealed one additional mutation. The mutations include two missense and one 4-base-pair deletion, and each mutation was found in patients with severe hemophilia. The polymorphism, located adjacent to the adenine branch site in intron 7, is useful for genetic prediction in some cases where the Bcl I and Xba I polymorphisms are uninformative. These results suggest that DNA amplification and denaturing gradient gel electrophoresis should be an excellent strategy for identifying mutations and polymorphisms in defined regions of the factor VIII gene and other large genes.

• OSTI ID: 6074714
• Journal Information: Proceedings of the National Academy of Sciences of the United States of America; (USA), Journal Name: Proceedings of the National Academy of Sciences of the United States of America; (USA) Vol. 87:6; ISSN PNASA; ISSN 0027-8424
• Country of Publication: United States
• Language: English

Related Subjects

550200* -- Biochemistry
59 BASIC BIOLOGICAL SCIENCES
ANIMALS
BLOOD COAGULATION FACTORS
CHROMOSOMES
COAGULANTS
DISEASES
DNA
DNA SEQUENCING
DRUGS
ELECTROPHORESIS
GENE AMPLIFICATION
GENE MUTATIONS
HEMATOLOGIC AGENTS
HEMIC DISEASES
HEMOPHILIA
HEREDITARY DISEASES
HETEROCHROMOSOMES
MAMMALS
MAN
MOLECULAR STRUCTURE
MUTATIONS
NUCLEIC ACIDS
ORGANIC COMPOUNDS
PRIMATES
PROTEINS
RFLPS
STRUCTURAL CHEMICAL ANALYSIS
VERTEBRATES
X CHROMOSOME