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

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.

我们通过减法和差异杂交的组合方法从富含视网膜的cDNA文库中克隆了几个新基因。通过鱼方法对这些候选基因进行染色体映射后，我们根据眼病的染色体图的信息选择了三个新基因并分析了它们。1。肌动蛋白（Myoc）与牛蛙的肌球蛋白和嗅觉素有同源性，位于感光细胞的纤毛根和基底体中。人们意识到，肌动蛋白与独立报道的蛋白质TIGR相同，该蛋白质TIGR是造成原发性开头青光眼的发病机理的。在小梁网细胞中表达了肌动蛋白。我们在大约100名日本青光眼患者中发现了MYOC基因中的3个突变。我们还隔离了肌动蛋白的小鼠同源物。2。包含富含亮氨酸的重复（ISLR）的免疫球蛋白超家族被认为是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很重要。北印迹分析表明，除大脑以外的各种组织中存在2.4 kb转录本。 ISLR映射到I 5q23-Q24.3染色体。视网膜特异性胺氧化酶（RAO）包含一个铜胺氧化酶的保守结构域，并将其映射到17q21染色体。 Rao在视网膜的神经节细胞层中表达，可能与GABA的合成有关。我们还分离了Rao.4的小鼠同源物。我们分离的新型候选基因之一被映射到2染色体。据报道，具有青光眼的大家族映射到该区域。我们正在分析新颖的克隆。