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AT Thliveris, DJ Zack, RW Nickells; Cloning and Characterization of a Novel cDNA that is Down-Regulated in Primate Experimental Glaucoma . Invest. Ophthalmol. Vis. Sci. 2002;43(13):4062.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose: To characterize a cDNA isolated by differential expression in control and glaucomatous monkey retinal cells. Methods: Differential display PCR (ddPCR) was used to screen first strand cDNA made from control and glaucomatous monkey retinas. Primers specific for one clone (4-6.7) were used for RT-PCR analysis of monkey tissue RNAs and different monkey cDNA libraries. Sib-selection PCR screening was used to isolate a longer clone of 4-6.7 from a control monkey cDNA library and a BAC clone from a human genomic library. Results: The sequence of 4-6.7 was homologous to a 240 bp sequence from human genomic clone RP11-279I21 (AL157936) corresponding to chromosome 9q22.2-31.1. PCR analysis indicated the 4-6.7 sequence was retina specific and found only in control samples, but not in retinas from either glaucomatous or optic nerve transected eyes. Sib-selection PCR screening of a monkey retina cDNA library yielded a rare 1.6 kB cDNA that contained the 4-6.7 ddPCR clone sequence. No obvious reading frame was detected in the 1.6 kb clone. The comparison between the monkey 1.6 kb cDNA and human genomic sequence is under investigation, but preliminary alignment suggests that the 6.7 clone originated from either genomic DNA or untranslated RNA. Conclusion: A novel cDNA that is down-regulated in glaucoma, has been isolated. The function of this cDNA remains a mystery.
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