摘要
目的建立基因MLXIPL、PMVK、TRIB3启动子区DNA甲基化检测的方法。方法用亚硫酸氢钠和对苯二酚处理外周血细胞基因组DNA标本,以修饰后的DNA标本为模板,每个基因分别用内外侧两套不同的引物对启动子区进行巢式扩增。PCR产物进一步克隆、测序。结果测序结果表明,MLXIPL、PMVK、TRIB3启动子区基因中的非甲基化的C碱基转变为T碱基,而CpG岛被甲基化的C碱基则不改变。结论成功地建立了MLXIPL、PMVK、TRIB3基因甲基化检测的方法,为探索基因启动子区甲基化与疾病的关系做出了铺垫。
Objective To establish time-efficient and sensitive method for detection of the methylation in MLXIPL, PMVK, TRIB3 genes promoter. Methods The two sets of primers were used to amplify each target DNA fragments by nest PCR combined touch-down PCR. The PCR products were further cloned and sequenced. Results The sequencing results showed that unmethylated cytosine residues （C） were transformed to uracil （U）, while methylated cytosine （mC） remained unchanged. Conclusion In the pre- sent study, methods for the detection of methylation states of MLXIPL, PMVK, TRIB3 genes promoters were successfully established, which provide a technique for exploring the relation between disease and the methylation of these genes.
出处
《中国实验诊断学》
北大核心
2010年第8期1166-1169,共4页
Chinese Journal of Laboratory Diagnosis
