Mutational Analysis of the Factor IX Gene in Eight Families with Hemophiaia B
Huling, Lenore P.
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Hemophilia B is an X-linked recessive blood coagulation disorder caused by a deficiency in the blood serine protease zymogen antihemophiliac factor IX, one of the intermediate steps in the complex cascade of reactions leading to the formation of a normal blood clot. Germline mutations in the gene that codes for factor IX limit the ability of the individual to form blood clots to varying degrees depending upon the degree to which the protein is rendered defective by the mutation. This project serves to delineate the causative mutations in eight hemophilia B patients in order to add to the already availiable database of published mutations. Nearly 3,500 bp of the factor IX gene, including eight exons, the 5' promoter region, and splice junctions were directly sequenced using a method known as genomic amplification with transcript sequencing (GAWTS). GAWTS is based on amplification by polymerase chain reaction (peR) and direct sequencing of single stranded RNA transcripts by the dideoxy chain termination method. Causative mutations were determined for seven of the eight patients sequenced. Using the same method, carrier testing was performed for female relatives wishing to be tested in order to determine if they were carriers of the deleterious allele. In two cases, the origin of the mutation was determined through carrier testing.