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H Xu, TS Acott, MK Wirtz; Identification and Mutation Screening of a Novel Candidate Gene in the GLC1F Region . Invest. Ophthalmol. Vis. Sci. 2002;43(13):1042.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose: A novel expressed sequence tag (EST) was identified localizing to the GLC1F region on chromosome 7 through random sequencing of over a thousand clones from a human trabecular meshwork (HTM) cDNA library. The purpose was to characterize the entire cDNA encoding this novel EST to allow mutation screening for GLC1F. Methods: Random sequencing of clones from a HTM cDNA library was used to identify this novel EST. Northern analysis was performed to determine the mRNA size. In silico cloning was conducted to retrieve the entire cDNA sequence from the Unigene database for related ESTs and align them using Seqencher program. The consensus sequence was blasted against genome database to obtain information on intron-exon boundary. Monoallelic mutation screening of this gene is being conducted on the family members whose glaucoma-causing gene was previously mapped to the GLC1F region by our group. Results: A novel EST was identified from the HTM cDNA library. Northern analysis showed that the message is expressed in eight different human tissues, including heart, brain, placenta, lung, liver, skeletal muscle, kidney, and pancreas. A single transcript of 2.9kb was observed. Approximately 2.9 kb of sequence was obtained through in silico cloning. Six exons comprising the entire coding sequence were identified from 6893 bp of genomic sequence. Monoallelic mutation screening of this novel gene in affected GLC1F members is being conducted . Conclusion: A novel gene has been identified localizing to the GLC1F region. It is expressed in the HTM and eight different human tissues. Based upon its chromosomal location, this novel gene is an ideal glaucoma candidate gene.
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