title: An alternative strategy for identification of type IIA VWD mutations.
abstract: A 306 base pair segment of the von Willebrand's factor (VWF) gene between nucleotides 4627 and 4932 was amplified by PCR using DNA from a type IIA VWD patient. The amplified DNA was cloned in the plasmid pCRII. Clones of the VWF gene and pseudogene were distinguished by their KpnI restriction patterns. A series of six VWF gene clones was sequenced. A single C to T point mutation at nucleotide 4789 resulting in the substitution of arginine 834 by tryptophan was identified in four clones. This mutation, which destroys a BstEII restriction site, was also detected in amplified DNA of affected relatives of the patient. The method described avoids difficulties associated with strategies employed previously for identification of VWD mutations.
authors.affiliation: Department of Haematology, Royal Victoria Hospital, Belfast, Northern Ireland.
authors.forename: P C
authors.initials: PC
authors.lastname: Winter
authors.forename: E E
authors.initials: EE
authors.lastname: Mayne
mesh_terms: D001483:Base Sequence; D004247:DNA / Q000737:chemistry; D006801:Humans; D008969:Molecular Sequence Data; D017354:Point Mutation / Q000235:genetics*; D016133:Polymerase Chain Reaction; D014842:von Willebrand Diseases / Q000235:genetics*; D014841:von Willebrand Factor / Q000235:genetics*
chemical_list: D014841:von Willebrand Factor; D004247:DNA