Mutation of the glucokinase gene has recently been identified as a cause of maturity-onset diabetes of the young (MODY), a subset of non-insulin-dependent diabetes mellitus (NIDDM). However, its role in the wide variety of NIDDM remains controversial due to conflicting reports of association studies, negative results of linkage studies and low prevalence of glucokinase mutations in the common variety of NIDDM. In this study, two (CA)n-microsatellite polymorphisms flanking both ends of the glucokinase gene, termed GCK1 and GCK2, were used to evaluate the role of glucokinase on NIDDM susceptibility of Taiwanese. For GCK1, three alleles (Z,Z+2 and Z+4 with a polymorphic information content index (PIC) of 0.53) and six genotypes were evident in 119 Taiwanese. When compared with control subjects, the NIDDM group had a much less frequency of the Z+2 allele (14.0% vs. 23.9%). In addition, the Z+2 allele was noted to have a marginal protective effect for NIDDM in Taiwanese with the odds ratio of 0.52 (95% confidence interval (C.I.) 0.26-1.03, P = 0.058). For GCK2, four alleles (0, 2, 4 and 6 with a PIC of 0.48) and seven genotypes were identified. There was no significant difference in allele frequency between NIDDM and control groups in the locus of GCK2. Our data were in agreement with reports from American Blacks, Mauritian Creoles, Asian Indians, Japanese and Finnish--that there is a positive association of GCK1 and a negative association of GCK2 with NIDDM. Furthermore, the Z+2 allele was a protective factor for NIDDM in Taiwanese.