CLONING RETINAL GENES LOCATED ON CHROMOSOME 11

遗传性眼病的基因编码已被家族绘制出来 对两个特定区域的研究和遗传连锁测试(11 Q13和 11p14/15.1)。这项建议的目的是定位 并克隆这些与视力受损有关的基因。这将是 通过生成包含以下内容的有序重叠克隆图来完成 第二染色体特异YAC(酵母人工染色体)克隆的交叉 通过连锁分析确定的含有疾病基因的区域。 将以1 Mb的间隔从YAC克隆开发CA重复，并使用 通过连锁分析鉴定与该病相邻的YAC克隆 精神错乱。基因选择和外显子捕获将直接应用于 从这些基因组克隆中确定编码区 包含这些基因的最小区域。得到的cdna克隆将是 测序以测试编码区的标准。候选cDNAs和 相应的基因组克隆将用于检查受影响的患者 单链构象多态(SSCP)和 变性梯度凝胶电泳法(DOGE)。分子和遗传学 每种疾病的异常情况将通过克隆和 描述导致这些视力障碍的基因。最终 基因的测序将使蛋白质的测定成为可能 由基因及其生理功能编码的结构。这一知识 将使人们对几种常见的视力障碍有新的见解。

Genes coding for inherited eye disorders have been mapped by family studies and genetic linkage tests to two specific regions (11 q13 and 11p14/15.1) on human chromosome 11. The aim of this proposal is to locate and clone these genes associated with impaired sight. This will be accomplished by generating an ordered overlapping clone map consisting of chromosome ii specific YAC (yeast artificial chromosome) clones across the regions identified by linkage analysis to harbor the disease genes. CA repeats will be developed from YAC clones at 1 Mb intervals and used to identify by linkage analysis those YAC clones flanking the disease loci. cDNA selection and exon trapping will be applied to directly identify coding regions from those genomic clones identified in the smallest region harboring the genes. Resulting cDNA clones will be sequenced to test for criteria of coding regions. Candidate cDNA and corresponding genomic clones will be used to examine affected patients for mutations by single strand conformational polymorphisms (SSCP) and denaturing gradient gel electrophoresis (DOGE). The molecular and genetic abnormality for each disorder will be described by cloning and characterizing the genes responsible for these sight disorders. Eventual sequencing of the genes will allow a determination of the protein structure coded by the gene and its physiologic function. This knowledge will allow new insights into several common sight disorders.