CLONING RETINAL GENES LOCATED ON CHROMOSOME 11

编码遗传性眼病的基因已按家族绘制 对两个特定区域（11 q13和 11p14/15.1）。本提案的目的是 克隆这些与视力受损相关的基因。这将是 通过生成一个有序的重叠克隆图来完成， 的染色体ii特异性YAC（酵母人工染色体）克隆， 通过连锁分析确定的携带疾病基因的区域。 CA重复序列将以1 Mb间隔从YAC克隆中产生并使用 通过连锁分析来鉴定那些位于疾病侧翼的YAC克隆 的位点cDNA选择和外显子捕获将直接应用于 鉴定来自那些基因组克隆的编码区， 最小的基因区域。将得到的cDNA克隆 测序以测试编码区的标准。候选cDNA和 相应的基因组克隆将用于检查受影响的患者， 单链构象多态性（SSCP）突变， 变性梯度凝胶电泳（DOGE）。分子和基因 将通过克隆描述每种疾病的异常， 描述了导致这些视力障碍的基因最终 基因测序将允许确定蛋白质 基因编码的结构及其生理功能。这些知识 将让我们对几种常见的视力障碍有新的认识。

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.