检测组织激肽释放酶基因寡核苷酸多态性方法的建立

A Method for Detecting of DNA Polymorphisms of Tissue Kallikrein Gene by ASO Blot Hybridization Technique

目的 :检测组织激肽释放酶启动子区寡核苷酸多态性 ,为开展基因诊断高血压研究奠定基础。方法 :人外周静脉抗凝血提取基因组DNA ,PCR扩增目标启动子片段 ,ASO(allele specificoligonucleotide)斑点杂交检测基因型 ,并与测序法比较了解启动子等位基因多态性。结果 :有 4例杂交结果阳性 ,与测序结果吻合。结论 :应用ASO斑点杂交技术检测该寡核苷酸多态性与测序法比较具有简便 ,快速 ,价廉 ,特异性高等优点 ,适于大规模人群筛查 。

s Objective: To detect DNA polymorphism in the 5' flanking region of the human tissue kallikrein gene promoter. Methods: Total DNA was extracted from whole blood,target promoter fragment was amplified by PCR, then detected by hybridization with the oligonucleotide probe and direct DNA sequencing. Results: The results of hybridization were coincident with those of DNA sequencing. 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.