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We have shown that a NfkappaBp50-dependent promoter (containing NkappaB binding sites) drives the expression of an L-2 (15% of pancreatic islet beta cell) promoter in INS-1E beta cell line, and that this promoter activity is dependent on NfkappaB DNA binding. We propose to further study the function of this promoter in the beta cell by deletion and mutational analysis of the L-2 promoter in both the INS-1E and beta cells, and by constructing a beta cell-specific promoter by linking the L-2 promoter to a beta-cell-specific promoter. We will also characterize the role of the NFkappaB family in regulation of insulin gene expression by a) identifying the proteins that bind to the NFkappaB site in the insulin promoter, b) analyzing the function of CRE and CCAAT box sites in the insulin promoter, and c) analyzing the effects of NFkappaB family members on insulin promoter activity. 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