MUTATIONS IN THE COL1A1 GENE OF TYPE I COLLAGEN

I型成骨细胞（OI）是遗传性脆性的最温和形式， 骨病 受影响个体的成纤维细胞产生大约一半的 预期数量的结构正常的I型胶原蛋白， 减少其组成链之一脯醛的合成。 PCR 扩增基因组DNA，然后进行单链构象 多态性电泳和变性梯度凝胶 已经使用DGGE技术来鉴定COL 1A 1中的突变， Ⅰ型胶原基因与轻度成骨不全的关系 一 个体被鉴定为具有单核苷酸取代， 5 ′供体剪接位点。 对该片段进行RT-PCR，以便cDNA可以 进行克隆和测序，以评估这种突变对剪接的影响。 此外，在内含子2区域检测到多态性 这在前面没有描述过。 表征这 polymorphism正在进行中。 未来的研究将包括分析 I型胶原的调节机制与OI患者的筛选 COL 1A 1基因第一内含子中的突变，其中调控元件 之前已经确认。 这些突变对基因的影响 将在成纤维细胞和骨细胞系中研究表达。

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 proal, 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 COLlA1 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 Ior mutations in the first intron of the COLlA1 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.