Abstract
The rat glucokinase (GK) gene containing the first exon was isolated and its 5′ flanking region was characterized by the bacterial chloramphenicol acetyltransferase (CAT) assay. A transient expression assay with a series of 5′ deletion constructs (−5.5k to −48) of GK-CAT fusion genes indicated that the 5′ flanking sequence up to nucleotide −87 was sufficient for promoter activity in adult rat hepatocytes, but its activity was much weaker than that of the SV40 enhancer/promoter. Similar promoter activity was also detected in dRLh-84 hepatoma cells, which do not express glucokinase. Insulin treatment caused no change in the CAT activity of hepatocytes transfected with the fusion genes. These results suggest that the 5′ flanking region of the glucokinase gene up to −5.5k does not contain enhancer elements responsible for tissue-specific expression or insulin regulation.
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