C57 BL/6J小鼠肝脏中mPer1基因CpG岛甲基化分析  

Analysis of methylation at CpG island of mPer1 gene in C57 BL/6J mouse liver

作　　者：刘姝[1] 蔡彦宁[1] 陈彪[1] 

机构地区：[1]首都医科大学宣武医院神经生物学室教育部神经变性病学重点实验室,北京100053

出　　处：《中国现代医学杂志》2009年第12期1813-1816,1820,共5页China Journal of Modern Medicine

基　　金：国家自然科学基金(No:30400148;30430280);科技部重点项目(No:2006CB500701);北京市科技新星计划(No:H020821400190);北京市自然基金(No:5063042)

摘　　要：目的建立分析C57BL/6J小鼠肝脏组织mPer1基因启动子区甲基化状态的亚硫氢酸修饰法,检测mPer1表达峰、谷对应的甲基化状态。方法利用实时定量PCR方法检测mPer1基因表达时相特点,并利用亚硫氢酸修饰法对小鼠肝脏组织mPer1基因启动子区域CpG岛和E-box甲基化情况进行了研究。结果C57BL/6J小鼠肝脏组织mPer1基因表达有显著波动性,表达高峰位于ZT13,低谷位于ZT01。序列分析显示mPer1基因启动子区含有CpG岛,并且该CpG岛覆盖与mPer1波动性表达密切相关的顺式元件E-box1和E-box2。亚硫氢酸修饰测序结果显示,mPer1基因启动子区CpG岛和E-box均呈低甲基化或非甲基化状态。不同时间点mPer1基因CpG岛甲基化频率无显著变化(P=0.458)。结论肝脏中mPer1基因启动子区CpG岛和E-box均为开放状态,可以与时钟相关转录因子结合。甲基化不参与调节mPer1基因的节律性表达。[Objective] To analyse the methylation of CpG dinucleotides in the promoter of mPerl gene in C57 BL/6J mouse liver. [Methods] Real-time PCR was employed to detect circadian expression of mPerl. Genomic DNA of mouse liver tissue were extracted and treated by bisulfite sodium and then sequenced. [Results] It indicated that mPerl oscillated around-the-clock with peak at ZT13 and through at ZT01. The 2 E-box in mPerl promoter region encompass 14 and 14 CpGs respectively, which were covered by a putative CpGs island. Both CpG island and E-boxes showed a low level of methylation, and even free from methylation. No time-dependent variations on DNA methylatian was observed in mPerl promoter region. [Conclusion] Methylation statuses of mPerl CpG island did not change over time in liver, suggesting that E-boxes are open to mBmall/mClock heterodimer, which is critical for mPerl daily oscillation. In addition, E-box methylation does not regulate the rhythmic expression of liver mPerl.

关 键 词：mPer1基因 E—box CPG岛 甲基化 实时定量PCR 

分 类 号：Q342[生物学—遗传学]