title: Non-phenotypic detection of osteopetrotic (op/op) mutation by using PCR-SSCP analysis.
abstract: Mice homozygous for recessive mutation osteopetrosis (op/op) on chromosome 3 provide a unique model to study the mechanism of haematopoiesis in conjunction with bone formation. Based on the DNA sequence data recently reported, we established a PCR-SSCP (polymerase chain reaction--single strand conformation polymorphism) assay which identifies an amplified fragment having an insertional point mutation present in macrophage colony-stimulating factor (M-CSF) gene of op/op mice. With this assay, three genotypes, op/op, +/op, and +/+ can be distinguished. Although heterozygous (+/op) and normal (+/+) mice could not be discriminated phenotypically, we could generate op/op mutant mice starting from a single heterozygous (+/op) mouse using only the PCR-SSCP aided screening method. This assay will permit introduction of the op mutant into any strain to generate a new animal model to study the cytokine network and haematopoiesis.
authors.affiliation: Department of Pathology, National Children's Medical Research Center Setagaya-ku, Tokyo, Japan.
authors.forename: H
authors.initials: H
authors.lastname: Kaneko
authors.forename: K
authors.initials: K
authors.lastname: Iizuka
authors.forename: T
authors.initials: T
authors.lastname: Morohashi
authors.forename: Y
authors.initials: Y
authors.lastname: Kokai
authors.forename: J
authors.initials: J
authors.lastname: Fujimoto
mesh_terms: D000818:Animals; D001483:Base Sequence; D004252:DNA Mutational Analysis / Q000379:methods*; D005260:Female; D005269:Femur / Q000473:pathology; D006579:Heterozygote; D007866:Leg / Q000000981:diagnostic imaging; D016173:Macrophage Colony-Stimulating Factor / Q000235:genetics*; D008297:Male; D051379:Mice; D008969:Molecular Sequence Data; D010022:Osteopetrosis / Q000235:genetics* / Q000473:pathology; D017354:Point Mutation; D016133:Polymerase Chain Reaction / Q000379:methods*; D018807:Polymorphism, Single-Stranded Conformational; D011859:Radiography
chemical_list: D016173:Macrophage Colony-Stimulating Factor