Human tissue kallikrein gene polymorphisms were identified in the promoter region by polymerase chain reaction (PCR) and DNA sequencing. One polymorphic region was identified between nucleotides -121 and -133 with respect to the transcription initiation site of the tissue kallikrein gene. Ten alleles with length and nucleotide sequence variations were detected among 108 unrelated Caucasians, African-Americans, and Asians. The polymorphisms show Hardy-Weinberg equilibrium. Allele-specific amplification and PCR analyses were used to detect the various forms of polymorphism. The promoter activity was analyzed in human embryonic kidney 293 cells by transient transfection assays. Sequential 5'-deletion analysis of the tissue kallikrein gene promoter revealed that the region from -144 to -98 is crucial for its promotor activity, while alleles D and H had significantly lower promoter activities than the other alleles in the -940/+10 deletion constructs. The high variability and the proximity to the tissue kallikrein gene render it suitable for application as a new tool in genetic studies for evaluation of the tissue kallikrein gene in the pathogenesis of human essential hypertension.