MOLECULAR GENETICS OF HEMOPHILIA A

血友病A是一种常见的血液凝固障碍，由于缺乏 凝血因子VIII。 第八因子的基因已被克隆， 特征和搜索已经开始了解的分子基础， 血友病A 在这个建议中，我们将描述所有的分子 在大约300名患有各种形式血友病A的患者中发现了缺陷。 突变 检测包括PCR产物的筛选（通过变性梯度凝胶 电泳或单链电泳或其它方法）和 突变体扩增DNA产物的核苷酸测序。 的突变 改变VIII因子蛋白重要区域的氨基酸， 分析它们对这种蛋白质功能的影响。 的 分析将涉及CRM阳性血浆中因子VIII的研究， 突变型cDNA瞬时表达后因子VIII的研究 哺乳动物细胞 拟议的工作将使人们更清楚地了解 1）突变的性质和产前起源，2）结构-功能 与第八因子蛋白的关系。

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.