MOLECULAR GENETICS OF HEMOPHILIA A

血友病A是一种常见的凝血障碍，由于缺乏 凝血因子VIII。因子VIII的基因已被克隆和 特征和研究已经开始了解的分子基础 血友病A。在这个提案中，我们将描述所有的分子。 约300例各种形式的血友病A突变患者的缺陷 检测包括对PCR产物的筛选(通过变性梯度凝胶 电泳法或单链电泳法或其他方法)和 突变扩增DNA产物的核苷酸测序。突变 改变氨基酸在重要区域的第VIII因子蛋白将是 分析了它们对这种蛋白质功能的影响。这个 分析将涉及对CRM阳性血浆中第VIII因子的研究和 突变型基因瞬时表达后凝血因子VIII的研究 哺乳动物细胞。拟议中的工作将提供更清晰的理解 1)突变的性质和先天起源；2)结构-功能 蛋白因子中的相互关系。

Hemophilia A is a common disorder of blood coagulation due to deficiency of clotting Factor VIII. The gene for factor VIII has been cloned and characterized and a search has begun to understand the molecular basis of hemophilia A. In this proposal, we will characterize all the molecular defects in about 300 patients with various forms of hemophilia A. Mutation detection includes screening of PCR products (by denaturing gradient gel electrophoresis or single stranded electrophoresis or other methods) and nucleotide sequencing of mutant amplified DNA products. Mutations that change amino acids in important areas of the factor VIII protein will be analyzed for their consequences on the function of this protein. The analysis will involve study of factor VIII in CRM positive plasmas and study of factor VIII after transient expression of mutant cDNAs into mammalian cells. The work proposed will provide a clearer understanding of: 1) nature of mutations and prenatal origin and 2) structure-function relationship in the factor VIII protein.