In this report, we describe transcriptional regulation of the human Gal beta 1,3 GalNAc alpha 2,3-sialyltransferase II (hST3Gal II) gene. The results of 5'-RACE showed that the forms of two mRNAs differed only in the 5'-untranslated region (Types 1 and 2). According to analysis of the genomic structure, the transcriptional regulation of Type 1 and Type 2 mRNA isoforms depended on the p1 and p2 promoters, respectively. Both the mRNA isoforms were detected in various human tissues except colon, skeletal muscle, and peripheral blood leukocytes by RT-PCR analysis. In colon tissue, the Type 2 mRNA was detected, however, Type 1 mRNA was not detected. To elucidate the molecular basis of hST3Gal II gene expression, we isolated and characterized the function of the genomic region of hST3Gal II containing the p1 and p2 promoters. The activity of p2 promoter is much higher than that of the p1 promoter in the colon adenocarcinoma cell line, COLO205. These results suggest that the hST3Gal II gene is expressed specifically by alternative promoter utilization and is regulated in a tissue-restricted fashion at the level of transcription.