GENETIC HETEROGENEITY: EHLERS DANLOS SYNDROMES VII & IV

胶原蛋白是一种组织特异性结构蛋白家族，其 生物合成异常可能影响MOST的发育和完整性 包括皮肤、关节和血管在内的器官(埃勒斯-丹洛斯综合征)；骨骼 (成骨不完善)；以及主动脉和心脏(马凡氏综合征)。那里 在任何类别的患者中都存在巨大的临床异质性， 反映了必须存在的分子异质性。遗传 胶原蛋白和其他细胞外基质成分的紊乱，如 弹性蛋白和纤维连接蛋白，提供了一个了解结构和 皮肤和结缔组织的功能关系。基因的克隆 这些蛋白质的DNA和基因以及聚合酶的发展 链式反应为分子方法提供了必要的工具 突变分析。 该项目将在选定的和 之前研究过的胶原蛋白生物合成障碍患者： Ehler-Danlos综合征和Buschke-Ollendorf综合征。蛋白质生物合成 SDS PAGE、肽图谱、Southern和Northern印迹 将被用来识别患者，进行更详细的分析。限制 片段长度多态(RFLP)将用于单倍型胶原 家系中的链等位基因以识别受影响的染色体(S)。DNA来自 疑似点突变的患者将使用RNAase A进行研究 保护分析是为了定位突变。这些突变将被测序 通过从成纤维细胞mRNA和DNA中合成第一链CDNA 聚合酶链式反应(PCR)扩增。通过使用不相等 低聚物的摩尔当量，单链DNA分子可以 双向扩增，用于突变的直接DNA测序。自.以来 胶原蛋白对传统的蛋白质测序具有抵抗力，这种分子 研究胶原蛋白生物合成异常的遗传学方法可能是 非常有用，提供有关结构和意义的信息 胶原蛋白在皮肤形态发生中的作用，并导致基因的发现 用于产前诊断的标记和可识别的突变 精神错乱。

Collagens are a family of tissue specific structural proteins, whose abnormal biosynthesis may affect the development and integrity of most organs including skin, joints, and vessels (Ehlers-Danlos Syndromes); bone (Osteogenesis Imperfects); and aorta and heart (Marfan's Syndrome). There is tremendous clinical heterogeneity among patients in any category, reflecting the molecular heterogeneity which must be present. Genetic disorders of collagen, and other extracellular matrix components such as elastin and fibronectin, provide an opportunity to understand structure and functional relationships of skin and connective tissue. The cloning of the cDNAs and genes for these proteins plus the development of the polymerase chain reaction provide essential tools for a molecular approach for analysis of mutations. This project will identify molecular genetic defects in selected and previously studied patients with disorders of collagen biosynthesis: Ehlers-Danlos Syndrome and Buschke-Ollendorf Syndrome. Protein biosynthesis SDS PAGE, peptide mapping, Southern and Northern blotting have been and will be used to identify patients for more detailed analysis. Restriction fragment length polymorphisms (RFLPS) will be used to haplotype collagen chain alleles in families to identify the affected chromosome(s). DNA from patients suspected point mutations will be studied using RNAase A protection assays to localize mutations. These mutations will be sequenced by performing 1st strand CDNA synthesis from fibroblast MRNA and amplification by the polymerase chain reaction (PCR). By using unequal molar equivalents of oligomers, single stranded DNA molecules can be amplified in both directions for direct DNA sequencing of mutations. Since collagens are resistant to conventional protein sequencing, a molecular genetic approach to the study of abnormal collagen biosynthesis may be extremely useful, provide information on the structure and significance of collagen in skin morphogenesis, and lead to the discovery of genetic markers and identifiable mutations for prenatal diagnosis of these disorders.