Cloning of retina-specific cDNAs and functional analysis

视网膜特异性 cDNA 的克隆和功能分析

基本信息

批准号：
09470384
负责人：
MASHIMA Yukihiko
金额：
$ 2.3万
依托单位：
Keio University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1997
资助国家：
日本
起止时间：
1997 至 1998
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09470384/
关键词：
retina gene cloning myocilin amine oxidase glaucoma mutation ISLR ミオシリン網膜アミンオキシダーゼ cDNA 染色体マッピング

项目摘要

We cloned several novel genes from retina-enriched cDNA library by the combined methods of subtractive and differential hybridization. After chromosomal mapping of these candidate genes by FISH method, we selected three novel genes according to the information of chromosomal mapping of eye diseases and analyzed them.1. Myocilin (MYOC) has homology to myosin and olfactomedin of bullfrog and is located in the ciliary rootlet and basal body of the connecting cilium of photoreceptor cell. It was realized that myocilin was identical to the independently reported protein TIGR which is responsible for pathogenesis of primary open angle glaucoma. Myocilin was expressed in the trabecular meshwork cells. We identified 3 mutations in the MYOC gene in about 100 Japanese glaucoma patients. We also isolated the mouse homologue of myocilin.2. Immunoglobulin superfamily containing leucine-rich repeat (ISLR) is considered to be a new member of the Ig superfamily whose domains are important for protein-protein interaction or cell adhesion. Northern blot analysis showed the presence of a 2.4 kb transcript in various tissues except brain. The ISLR was mapped to chromosome I 5q23-q24.3. Retina-specific amine oxidase (RAO) contained a conserved domain of copper amine oxidase and was mapped to chromosome 17q21. RAO was expressed in the ganglion cell layers of the retina and may be associated with the synthesis of GABA.We also isolated the mouse homologue of RAO.4. One of the novel candidate genes we have isolated was mapped to chromosome 2. A large family with glaucoma is reported to mapped to this region. We are analyzing the novel clone.

我们通过消减和差异杂交相结合的方法从富含视网膜的cDNA文库中克隆了几个新基因。通过FISH方法对这些候选基因进行染色体定位后，根据眼部疾病染色体定位信息筛选出3个新基因并进行分析。 1．肌球蛋白（MYOC）与牛蛙的肌球蛋白和嗅球素具有同源性，位于感光细胞的睫状根和连接纤毛的基体中。人们认识到肌纤蛋白与独立报道的蛋白质 TIGR 相同，TIGR 负责原发性开角型青光眼的发病机制。肌纤蛋白在小梁网细胞中表达。我们在大约 100 名日本青光眼患者的 MYOC 基因中发现了 3 个突变。我们还分离出了myocilin的小鼠同源物。2。含有富含亮氨酸重复序列的免疫球蛋白超家族（ISLR）被认为是 Ig 超家族的新成员，其结构域对于蛋白质-蛋白质相互作用或细胞粘附非常重要。 Northern 印迹分析显示除脑外的多种组织中都存在 2.4 kb 转录物。 ISLR 被定位到 I 号染色体 5q23-q24.3。视网膜特异性胺氧化酶 (RAO) 包含铜胺氧化酶的保守结构域，被定位到染色体 17q21。 RAO在视网膜神经节细胞层表达，可能与GABA的合成有关。我们还分离出了RAO的小鼠同源物。4。我们分离出的新候选基因之一被映射到 2 号染色体。据报道，一个患有青光眼的大家族被映射到该区域。我们正在分析新的克隆。