MUTATIONS IN THE COL1A1 GENE OF TYPE I COLLAGEN

I 型胶原蛋白 COL1A1 基因突变

• 批准号: 3753477
• 负责人: REBECCA L SLAYTON
• 金额: --
• 依托单位: UNIVERSITY OF IOWA
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3753477
• 关键词: collagen; fibroblasts; gel electrophoresis; gene expression; gene mutation; genetic mapping; genetic polymorphism; human genetic material tag; molecular cloning; nucleic acid sequence; osteogenesis imperfecta; polymerase chain reaction; protein biosynthesis; structural genes

Osteogenesis imperfecta (OI) type I is the mildest form of inherited brittle bone disease. Fibroblasts from affected individuals produce about half the expected amount of structurally normal type I collagen as a result of decreased synthesis of pro`1, one of its constituent chains. PCR amplification of genomic DNA followed by single-stranded conformational polymorphism electrophoresis (SSCP) and denaturing gradient gel electrophoresis (DGGE) has been used to identify mutations in the COLIAI gene of type I collagen in patients with mild Osteogenesis imperfecta. One individual was identified with a single nucleotide substitution which alters the 5' donor splice site. RT-PCR was done on this fragment so that cDNA can be cloned and sequenced to evaluate the effect of this mutation on splicing. In addition, a polymorphism has been detected in the region of intron 2 which has not been previously described. Characterization of this polymorphism is in progress. Future studies will involve analysis of the regulatory mechanisms of type I collagen and screening OI patients for mutations in the first intron of the COLIAI gene, where regulatory elements have been previously identified. The effect of these mutations on gene expression will be studied in fibroblast and osseous cell lines.

I型骨生成障碍（OI）是遗传性骨形成障碍的最温和形式。 脆骨病 受影响个体的成纤维细胞产生 约为预期结构正常的I型胶原蛋白量的一半， 这是其组成链之一pro 1合成减少的结果。 PCR扩增基因组DNA，然后进行单链 构象多态性电泳（SSCP）和变性 梯度凝胶电泳（DGGE）已被用于鉴定突变 轻度成骨患者I型胶原的COLIAI基因 你好 其中一个人被鉴定出有一个核苷酸 取代，其改变5'供体剪接位点。 RT-PCR在 这样cDNA就可以被克隆和测序， 这种突变对剪接的影响。 此外，多态性具有 在内含子2的区域中检测到了以前没有被检测到的 介绍了 这种多态性的表征正在进行中。 未来的研究将包括分析的调控机制， I型胶原和筛选OI患者的第一个突变， COLIAI基因的内含子，其中调控元件已被 以前识别的。 这些突变对基因表达的影响 将在成纤维细胞和骨细胞系中进行研究。