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X.C. Zhao, R.W. Yee; Corneal Expression of Candidate Genes for chromosome 10-linked Thiel-Behnke Corneal Dystrophy (CDB2) . Invest. Ophthalmol. Vis. Sci. 2003;44(13):3866.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose: We previously mapped Thiel-Behnke corneal dystrophy to chromosome 10q23-q24 (Yee et al. 1997, Genomics 46:152-154). Our goal is to clone this disease gene by positional cloning and candidate gene approaches. Several cloned genes in which mutations cause other forms of corneal dystrophy are expressed in the cornea. Therefore, we hypothesize that the gene responsible for this disease is expressed in the cornea. As a first step toward to cloning of this disease gene, we want to identify candidate genes by database search and analysis and prioritize candidate genes by determining their expression in human corneal tissues. Methods: GenBank, UniGene, and other molecular databases were searched for candidate gene identification. PCR primers were then designed for each of the candidate genes using PrimerSelect of DNA STAR programs. cDNA was PCR-amplified from a human corneal cDNA library (Stratagene) using T3 and T7 primers and purified using Qiagen PCR purification kit. This corneal cDNA was then used to amplify each of the selected candidate genes. The amplification pattern of each candidate gene was compared to genomic DNA control. Results: The database search identified a total of 74 candidate genes within the CDB2 locus interval. Twenty-five of them are known genes, and the rest are either expressed sequences or predicted genes by gene-finding programs. Analysis of these known genes through database search found that 14 of the 25 genes were expressed in the human eyes. Gene expression analysis confirmed the expression these 14 genes in the human cornea and identified additional 6 genes whose expression in human cornea was not known. Among 49 ESTs and predicted genes, 36 were found to express in the human cornea. A total of 17 candidate genes are not expressed in the human cornea and can be considered lower priority candidates genes in candidate gene analysis and mutation screening. Conclusions: The human genome project and associated databases greatly facilitate identification and cloning of human disease genes. Corneal-expressed genes identified within the CDB2 locus are higher priority candidates for the disease gene in mutation screening and analysis. Gene expression can be a valuable tool in prioritizing candidate genes in a positional cloning and candidate gene cloning project.
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