【摘要】 目的 :建立聚合酶链反应—限制性片段长度多态性 (PCR- RFL P)分析结合 DNA直接银染技术检测载脂蛋白 E基因多态性。方法 :选择江苏地区汉族健康人 16 8例及冠心病 6 3例 ,PCR扩增 Apo E基因第 4外显子包含编码第 112位和第 15 8位氨基酸残基的基因序列 ,cfo I限制性内切酶酶切后电泳 ,银染色后分析 Apo E基因型。结果 :汉族健康人群及冠心病人群 Apo E基因型频率分别为 :ε2 /2 0 .6 % ,0 ;ε2 /3 11.9% ,14.3% ;ε2 /4 1.2 % ,4.8% ;ε3/4 10 .7% ,17.5 % ;ε3/3 75 .0 % ,6 1.9% ;ε4/4 0 .6 % ,1.6 % ;Apo E等位基因频率分别为 :ε2 7.14% ,9.5 2 % ;ε386 .31% ,77.78% ;ε46 .5 5 % ,12 .70 %。结论 :PCR- RFL P方法快速、简便、准确性和可靠性高 ,适合普通实验室开展及大规模研究更多还原

【Abstract】 Objective:To establish polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP)combining silver stainning to detect Apolipoprotein E gene polymorphism.Methods: 168 cases of normal Chinese of Han nationality in Jiangsu area and 63 cases of patients with coronary heart diseases were collected. Exon 4 of Apolipoprotein E fragment coding for 112 and 158 amino acid residues was amplified by PCR, and the amplicons was digested by CfoI endonuclease ,after electrophoresis and silver staining,the specific CfoI fragments of different sizes were visualized.Results:The frequencies of Apolipoprotein E genotypes in normal subjects and patients with coronary heart disease separately were:ε2/2 0.60%,0;ε2/3 11.90%,14.29%;ε2/4 1.20%,4.76%;ε3/4 10.70%,17.46%;ε3/3 75.00%,61.90%;ε4/4 0.60%,1.59%; the frequencies of Apolipoprotein E alleles separatedly were:ε2 7.14%,9.52%;ε3 86.31%,77.78%;ε4 6.55%,12.70%.Conclusion:PCR-RFLP is a rapid and simple method for obtaining accurate and reliable result.更多还原