title: Molecular and genetic analysis of a compound heterozygote for dysprothrombinemia of prothrombin Tokushima and hypoprothrombinemia.
abstract: The molecular and genetic basis of a compound heterozygote for dys- and hypoprothrombinemia was analyzed. Abnormal nucleotide sequences of the human prothrombin gene were screened by PCR-single-strand conformation polymorphism (PCR-SSCP) with endonuclease digestion and mutated primer-mediated PCR-RFLP. A single nucleotide substitution responsible for dysprothrombinemia of prothrombin Tokushima was detected, as were three polymorphisms. The mutation for hypoprothrombinemia was detected by PCR-single-strand conformation polymorphism (PCR-SSCP) with endonuclease digestion in exon 6, near MboII-RFLP and NcoI-RFLP. Sequencing of PCR-amplified genomic DNA revealed a single base insertion of thymine (T) at position 4177. The resulting frameshift mutation caused both an altered amino acid sequence from codon 114 and a premature termination codon (i.e., TGA) at codon 174 in exon 7. Because exon 7 encodes the kringle 2 domain preceding the thrombin sequence, this frameshift leads to the null prothrombin phenotype. The inheritance of the hypoprothrombinemia gene from the father to the proband was proved by PCR-SSCP with endonuclease digestion and mutated primer-mediated PCR-RFLP.
authors.affiliation: Otsuka Department of Clinical and Molecular Nutrition, University of Tokushima, Japan.
authors.forename: H
authors.initials: H
authors.lastname: Iwahana
authors.forename: K
authors.initials: K
authors.lastname: Yoshimoto
authors.forename: T
authors.initials: T
authors.lastname: Shigekiyo
authors.forename: A
authors.initials: A
authors.lastname: Shirakami
authors.forename: S
authors.initials: S
authors.lastname: Saito
authors.forename: M
authors.initials: M
authors.lastname: Itakura
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D001778:Blood Coagulation Disorders / Q000150:complications / Q000235:genetics*; D002648:Child; D004247:DNA; D004252:DNA Mutational Analysis; D004720:Endonucleases; D005260:Female; D016368:Frameshift Mutation; D006580:Genetic Carrier Screening; D006801:Humans; D007020:Hypoprothrombinemias / Q000150:complications / Q000235:genetics*; D008297:Male; D008969:Molecular Sequence Data; D010375:Pedigree; D016133:Polymerase Chain Reaction; D011516:Prothrombin / Q000235:genetics*; D013941:Thymine
chemical_list: C051306:prothrombin Tokushima; D011516:Prothrombin; D004247:DNA; D004720:Endonucleases; D013941:Thymine