A Rapid Analysis of Human Paraoxonase Gene Cluster Polymorphism

 

A Rapid Analysis of Human Paraoxonase Gene Cluster Polymorphism

人芳香二烷基磷酸酯酶基因丛基因多态性快速分析法

 

摘要 ：目的　建立一种人芳香二烷基磷酸酯酶(PON)基因丛等位基因快速分型法。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PON1-192、PON1-55和PON2-31l位密码子的DNA片段，当等位基因为PON1-192R/PON1-55L/PON2-31lS时，PCR 扩增产物中均被引入唯一的限制性内切酶HinfⅠ的识别位点G(上标 +)ANTC。PCR 扩增产物经酶消化，聚丙烯酰胺凝胶电泳后，相互分开的不同组合的片段，确认为不同的基因型组合，同时对3个位点进行基因分型。结果　在检测的80例健康个体中，等位基因频率为PON1-192:Q46.9%.R53.1%:PON1-55:L95.6%，M4.4%；PON2-31l:S78.8%，C21.2%。结论　此方法快速分析3个位点的基因多态性，省时省力、节约经费，便于3个位点之间的连锁分析，具有推广价值。

Objective To establish an assay to identify paraoxonase gene cluster polymorphism. Methods Based on multiplex-PCR -RELP, we used mismatch primers that introduce an unique recognition site (Hinf Ⅰ) in PCR product in the presence of the R allele of PON1-192, L allele of PON1-55 and the S allele of PON-311 and then we simultaneously identified PON cluster polymorphism with electrophoretic band pattern specific for the variable combination through subsequent restriction analysis. Results The result shows that in 80 healthy individuals, the allelic frequency of PON1-192:Q46.9%, R 53.1%; PON1-55:L95.6% M4.4%; PON2-311:S78.8% C21.2% respectively. Conclusion This assay is an easy, economical and time-saving one characterized by detecting three polymorphism simultaneously.

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