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目的:为调查海南省老年性痴呆患者载脂蛋白E(Apo E)基因频率,建立一种简便、快速、准确的人载脂蛋白E基因型检测方法。方法:应用碘化钠法提取模板DNA,聚合酶链反应(PCR)特异性扩增Apo E基因含112 位和158 位氨基酸的基因序列,扩增产物用限制内切酶Hha I酶切,聚丙烯酰胺凝胶电泳后,观察酶切位点的限制性片段长度多态性(RFIP)图谱。结果:碘化钠法提取模板DNA 与经典法无差异;运用PCR- RFIP法检测30 名健康人Apo E基因型,实验成功。结论:该方法简单、快速、准确,适合于一般实验室开展Apo E基因型调查
Objective: To establish a simple, rapid and accurate method for detecting human apolipoprotein E genotypes in order to investigate the frequency of Apo E gene in patients with senile dementia in Hainan Province. Methods: The template DNA was extracted by sodium iodide method. The gene sequences of 112 and 158 amino acids of Apo E gene were amplified by polymerase chain reaction (PCR). The amplified product was digested with restriction endonuclease Hha I and poly After acrylamide gel electrophoresis, restriction fragment length polymorphism (RFIP) patterns were observed. Results: There was no difference between the template DNA extracted by sodium iodide and the classical method. PCR-RFIP was used to detect the Apo E genotype of 30 healthy people. The experiment was successful. Conclusion: The method is simple, rapid and accurate and suitable for general laboratory investigation of Apo E genotype