人蛋氨酸合成酶还原酶A66G基因突变频率的检测

目的 建立一种简便、准确、实用的人蛋氨酸合成酶还原酶(MSR)A66G基因突变频率的检测方法,并了解汉族人MSR基因的分布特点。方法 应用聚合酶链反应(PCR)特异性扩增蛋氨酸合成酶还原酶(MSR)A66G基因序列,扩增产物用限制性内切酶NdeI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(RFLP)图谱。结果 运用PCR-RFLP法检测了150名汉族人蛋氨酸合成酶还原酶(MSR)A66G基因点突变,其中野生型纯合子频率为25.33％,杂合子频率为52.00％,突变型纯合子频率为22.67％。突变等位基因频率为0.4867。结论 该方法简便、快速、准确,适合于一般实验室检测及大规模的人群调查,汉族人MSR基因的分布与其他地区人群的分布无明显差异。

Detection of mutation frequencies in the methionine synthase reductase gene with PCR-RFLP in health individuals

Objective To establish a simple, accurate and practical method for methionine synthase reductase (MRS) genotyping and to identify the distribution feature of MSR gene in the Hans nationality in Chinese. Method Methionine synthase reductase gene sequences were amplified by using polymerase chain reaction (PCR). The PCR products were digested with NdeI and subjected to electrophoresis on polyacrylamide gels. The patterns of restriction fragment length polymorphism(RFLP) of MRS gene were distinguished. Results 150 healthy individuals were investigated with this PCR-RFIP method. The prevalence of genotype for the normal homozygote, the normal/mutant heterozygote and the mutant homozygote was 25.33% ,52.00% and 22.67% respectively. The frequency of the mutant allele in the subjects was 0.4867. Conclusion The PCR-RFIP method was considered as a rapid and simple technique for obtaining accurate results and also suitable for routine laboratories and large-scale population studies,MSR genes distribution in the Hans in Chinese is undifferent from those in other areas.