机构地区: 武汉大学中南医院
出 处: 《武汉大学学报(医学版)》 2005年第1期1-4,共4页
摘 要: 目的 :建立一种对PON1 192和PON1 5 5位密码子等位基因同步分型的方法。方法 :在Multiplex PCR RELP基础上 ,通过引物设计错配 ,在一体系中同时扩增分别含密码子PON1 192和PON1 5 5的DNA片段 ,当等位基因为PON1 192R/PON1 5 5L时 ,其PCR扩增产物均被引入唯一的限制性内切酶HinfⅠ的识别位点G ANTC。PCR产物经酶消化 ,聚丙烯酰胺凝胶电泳后 ,根据不同组合片段 ,确认为不同的基因型 ,从而同时对两个位点进行基因分型。结果 :在检测的 80例健康个体中PON1 192 :Q 4 6 .9% ,R 5 3.1% ;PON1 5 5 :L 95 .6 % ,M 4 .4 % .结论 :此方法同时可分析基因两个位点的多态性 ,省时省力 ,方法简便 ,便于两个位点以及与疾病之间的连锁分析 ,很有推广价值 . Objective: To find an easy method to identify paraoxonase-1 gene cluster polymorphism simultaneously. Methods: With multiplex-PCR-RFLP, mismatching primers that introduce a unique recognition site (HinfⅠ) in the PCR product in case of presence of the R allele of PON1-192 and of the L allele of PON1-55 were used and then the PON1 polymorphism with an electrophoretic band pattern which was specific for the variable combination was identified through subsequent restriction analysis. Results: In 80 healthy individuals, the allelic frequency of PON1-192 were: Q 46.9% and R 53.1%; and that of PON1-55 were: L 95.6% and M 4.4%. Conclusion: This assay is an easy,economical and time-saving technique by detecting three polymorphism simultaneously.
关 键 词: 芳香二烷基磷酸酶 多重聚合酶链反应 基因多态性 动脉粥样硬化
领 域: [生物学]