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A.R. Djalilian, D.M. McGaughey, S. Patel, J.A. Segre; Characterization of the Promoter Region of Connexin 26 and Its Regulation by Transcription Factors . Invest. Ophthalmol. Vis. Sci. 2005;46(13):2113.
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Purpose: The expression of connexins (gap junction proteins) are spatially and temporally controlled in the corneal epithelium during normal and wound healing conditions. This study was performed to characterize the promoter region of connexin 26 and identify transcription factors that may regulate its expression. Methods: Multiple primers and RT–PCR were used to localize the transcription start site for mouse connexin 26 more precisely. The conserved sequences upstream of the human and mouse connexin 26 were identified using the Pipmaker program. Potential transcription factor binding sites were identified using TRANSFAC program. The regulation of the mouse connexin 26 promoter by transcription factors was assessed by a luciferase reporter assay following transient transfection into a mouse keratinocyte cell line. Results: The connexin 26 first exon was found to begin at least 250 bp upstream to that previously reported in Refseq. Besides the coding sequence, conserved sequences were identified within the intron regions and up to 6 kb upstream of the transcription start site. Both a 250 bp and 1 kb fragment immediately upstream of the first exon were found to have promoter activity. A transcription factor was identified with repressor activity on both the 250 bp and 1kb presumed connexin 26 promoter. Conclusions: This study demonstrates the use of bio–informatics tools for promoter analysis and provides insight into the regulation of the connexin 26 gene and its role in epithelial differentiation.
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