Gene expression analysis of human corneal and conjunctival epithelium by random cDNA sequencing

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We studied the composition of mRNA in human corneal epithelium by the random cDNA sequencing. The expression profile of active genes in human corneal epithelium showed that the most adundant transcripts represented those of genes for apolipoprotein J (apoJ), calcyclin, alpha-enolase, human adipose mRNA and a novel gene.The expression of both mRNA and protein of the most abundant trascripts, apoJ was investigated in corneal and conjunctival epithelium, as well as other mucosal epithelia (vagina, esophagus, stomach, small intestine, and colon) and epidermis. ApoJ mRNA transcripts was detected in corneal, conjunctival, vaginal and esophageal epithelium, but not in epidermal keratinocytes. ApoJ mRNA localized in all layrs of corneal epithelium, prominently in basal layr. ApoJ protein localized in the apical cell layrs of corneal and conjunctival epithelium, as well as various mucosal epithelia including vagina, esophagus stomach, small intestine and colon, but not in epidermis. These results indicated that apoJ may play a physiologically important role at the tear-ocular surface interface.We cloned the full lenght cDNA of the novel gene. The cDNA contained ORF highly homologous to mouse keratin 12, indicating that this gene is human cornea-specific keratin K12.Regarding the cDNA library construction of conjunctival epithelial cells, we are studying the sample collection method without contamination by blood cells or subepithelial cells.