Cloning of a Novel Gene from the Subtractive Retinal cDNA Library following Optic Nerve Injury

Abstract

Abstract: : Purpose:To clone and characterize a novel gene which was found to be up-regulated in the retina following optic nerve injury. Methods:Up-regulation of this novel gene was found in the subtractive hybridization and verified by RT-PCR. The mRNA and peptide sequences of this novel gene was determined using RT-PCR, TA cloning, sequencing, and bioinformatics. RNA expression profile was examined in adult mice tissue using Northern blot. The protein expression of this novel gene was investigated by immunostaining and Western blot. Results:Sequencing and bioinformatics revealed this novel gene to be a 5-kb transcript, which was confirmed on Northern blot. Predicted protein sequence and Western blot show a 65 kD peptide with a FERM domain. The immunostaining demonstrated that this novel gene is expressed on the retinal ganglion layer and implicated a cell-cell contact function of this gene. Conclusions:A novel gene was cloned and characterized from a subtractive retinal cDNA library following optic nerve injury.

Keywords: regeneration • gene/expression • ganglion cells