Song Q, Chao J, Chao L
Department of Biochemistry and Molecular Biology, Medical University of South Carolina, Charleston 29425, USA.
Hum Genet. 1997 Jun;99(6):727-34. doi: 10.1007/pl00008703.
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.
通过聚合酶链反应（PCR）和DNA测序在启动子区域鉴定了人组织激肽释放酶基因多态性。相对于组织激肽释放酶基因的转录起始位点，在核苷酸-121和-133之间鉴定出一个多态性区域。在108名无亲缘关系的高加索人、非裔美国人和亚洲人中检测到10个具有长度和核苷酸序列变异的等位基因。这些多态性呈现哈迪-温伯格平衡。采用等位基因特异性扩增和PCR分析来检测各种形式的多态性。通过瞬时转染实验在人胚肾293细胞中分析启动子活性。组织激肽释放酶基因启动子的5'-连续缺失分析表明，-144至-98区域对其启动子活性至关重要，而在-940/+10缺失构建体中，等位基因D和H的启动子活性明显低于其他等位基因。高度变异性以及与组织激肽释放酶基因的接近性使其适合作为一种新工具应用于基因研究，以评估组织激肽释放酶基因在人类原发性高血压发病机制中的作用。
