MOLECULAR GENETIC ANALYSIS OF LOWE'S SYNDROME

眼脑肾综合征（OCRL）是一种X连锁的先天性缺陷， 病因不明的代谢。 受影响的男性智力迟钝， 先天性白内障和肾小管功能障碍。 OCRL的位点有 通过与限制性片段长度的连接定位到Xq 25区域 在Xq 24 -26的多态性（RFLPs）和女性OCRL的发生， 具有在Xq 25处具有断裂点的X/3易位。 酵母人工 在大肠杆菌中分离了含有100-120 kb人插入片段的YAC染色体。 另一个，合作实验室，并提供给这个实验室。 这个YAC，RS 88，似乎包含来自 在OCRL女性中，X3易位中存在断点。 使用 来自RS 88内的序列，λ或λ中的重叠基因组序列， 粘粒载体将被分离，直到粘粒载体周围的整个区域被分离。 断点已克隆。 这个地区的所有DNA都将被检测 它是否包含候选基因的外显子，基于（i）保守性， 人与其他哺乳动物之间的核酸同源性，（ii）人与其他哺乳动物之间的核酸同源性的表达， 在透镜、肾、脑和成纤维细胞中的序列，（所有靶组织 OCRL），（iii）证明这些突变，缺失或其他， OCRL先证者的推定外显子序列。 当候选基因 识别， （一） 其整个mRNA序列和基因组结构将被表征。 （二） 导致疾病的突变将由 测序DNA中基因的外显子和内含子-外显子边界， 患者 (3)将产生针对基因产物的抗体， 表达基因的组织和细胞， 基因产物的亚细胞定位。 (4)负责控制基因启动子和增强子序列 表达将被识别。 长期的目标是确定人类智力基因 通过其位置而不是其功能， 来表征基因的表达和突变 最终的目标是了解 基因产物是什么以及基因缺陷如何导致疾病。

The oculocerebrorenal syndrome of Low (OCRL) is an X-linked inborn error of metabolism of unknown etiology. Affected males are retarded and have congenital cataracts and renal tubular dysfunction. The locus for OCRL has been mapped to the Xq25 region by linkage to restriction fragment length polymorphisms (RFLPs) in Xq24-26 and by the occurrence of OCRL in a female with an X/3 translocation with breakpoint at Xq25. A yeast artificial chromosome (YAC) containing a 100-120 kb human insert was isolated in another, collaborating laboratory and made available to this laboratory. This YAC, RS88, appears to contain sequences from either side of the breakpoint in the X;3 translocation in the female with OCRL. Using sequences from within RS88, overlapping genomic sequences in lambda or cosmid vectors will be isolated until the entire region around the breakpoint have been cloned. All DNA from this region will be tested for whether it contains exons of a candidate gene based on (i) conservation of nucleic acid homology between man and other mammals, (ii) expression of the sequence in lens, kidney, m brain and fibroblasts, (all target tissues for OCRL), (iii) demonstration of mutation, deletional or otherwise, of these putative exonic sequences in OCRL probands. When a candidate gene is identified, (1) its entire mRNA sequence and genomic structure will be characterized. (2) the mutations responsible for the disease will be determining by sequencing the exons and intron-exon boundaries of the gene in the DNA of patients (3) antibodies to the gene product will be raised and used to determine the tissue(s) in which the gene is expressed and the cellular and subcellular localization of the gene product. (4) the promoter and enhancer sequences responsible for control of gene expression will be identified. The long-term objective is to identify the gene for a human mental retardation syndrome through its location rather than its function and then to characterize the gene and in expression as well as the mutations responsible for the disease in man. The ultimate goal is to understand what the gene product is and how defects in the gene produce disease.