A Method For Detecting Of DNA Polymorphisms Of Tissue Kallikrein Gene

Objective One single nucleotide polymorphic region was identified between nucleotides -121and-133 with respect to the transcription initiation site of the human tissue kallikrein gene. Ten alleles with length and nucleotide sequence variations were detected. In this study, we will establish an efficient detective method to study the SNP in the 5'-flanking region of the human tissue kallikrein gene promoter, and detect the distribution frequency of the SNP in Chinese Hans, then determine whether it is associated with NIDDM.Method Total DNA was extracted from whole blood, target promoter fragment was amplified by PCR, then detected byhybridization with the oligonucleotide probe and direct DNA sequencing.Results The results of hybridization were coincident with those of DNA sequencing. Only A, B, H, K alleles were detectedin Chinese Hans. There was a statistical difference (P<0.05) of each allele frequency between NIDDMs and controls.Conclusion ASO(allele-specific oligonucleotide) blot hybridization technique is more economical than DNA sequencing to be used to detect the 5'-flanking polymorphism of tissue kallikrein gene promoter. The identified gene variants are associated with NIDDM.