The 615-bp 5' flanking region of the human TNF-alpha/cachectin gene was isolated and ligated to the luciferase reporter gene. In addition, a series of truncated promoter constructs was generated by exonuclease III digestion. The promoter activity of these constructs was studied in a transient transfection system using the TNF-alpha-producing U937 cell line. Full-length and truncated TNF promoter constructions extending from -615 to -95 bp relative to the transcription start site (TSS) could be induced by phorbol esters. A construct truncated to within 36 bp of the TSS (and within 11 bp of the TATAA box) was inactive. Therefore, the phorbol ester responsive is localized in the TNF/cachectin promoter to a relatively short region proximal to the TATAA box.
Genetic analysis of the human tumor necrosis factor alpha/cachectin promoter region in a macrophage cell line.
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J S Economou, K Rhoades, R Essner, W H McBride, J C Gasson, D L Morton; Genetic analysis of the human tumor necrosis factor alpha/cachectin promoter region in a macrophage cell line.. J Exp Med 1 July 1989; 170 (1): 321–326. doi: https://doi.org/10.1084/jem.170.1.321
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