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Carrier Detection of Ovine Hemophilia A Using an RFLP Marker, and Mapping of the Factor VIII Gene on the Ovine X-Chromosome

Abstract

Ovine hemophilia A is an X-linked recessive bleeding disorder. For diagnostic purposes, restriction fragment length polymorphism (RFLP) analysis in the region of the factor VIII (F-VIII) gene was carried out using human F-VIII gene probes. The probe St14, Known to detect a highly polymorphic region that is closely linked to the F-VIII gene in humans, hybridized nonspecifcally with DNA from sheep. Searching for Intragenic RFLPs, the entire 9.0-kb coding sequence of the human F-VIII was used as a probe. Using the 1.8-kb Sstl/Kpnl F-VIII cDNA probe for hybridization, an Mspl-RFLP with allelic bands of 5.8 Kb (A1) and 4.2 kb (A2) was detected. A1 was in linkage phase with the mutated allele responsible for hemophilia A. The F-VIII locus in the sheep genome was assigned to the long arm of the X-chromosome in the region Xq24-q33, Using In situ hybridization with a 3-Kb human F-VIII cDNA probe to QFQ banded sheep metaphase chromosome

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