摘要
利用实时定量PCR方法检测胸腺和睾丸组织中mPer1基因表达时相特点,采用亚硫氢酸修饰法对两种组织两个时间点该基因2个启动子区域5个E-box CpGs甲基化情况进行研究.结果显示:C57 BL/6J小鼠胸腺和睾丸组织中mPer1基因表达没有显著波动性,mPer1基因启动子区CpGs呈低甲基化状态(<10%).两种组织中E3和E4甲基化频率存在显著差异(P<0.01).在睾丸中,不同时间点E3和E4甲基化频率有显著变化(P<0.01).说明在睾丸中,mPer1启动子E-box3的甲基化状态能够随时间波动.但在胸腺和睾丸中,甲基化并不是mPer1基因调节的主要方式.
Objectives： To investigated whether CpG methylation contributes to the atypical roPer1 transcription in testis and thymus .Methods： Real time PCR was employed to detect circadian expression of mPerl gene. Five E-box-containing amplicons were examined in mouse testis and thymus at both early-loght （Zeitgeber time 01, ZT01 ） and early-dark （ZT13） using bisulfite genomic sequencing. Results： E-boxes were sparsely methylated in testis and thymus at both times. Significant different of E3 and E4 were found between thymus and testits. Significant increases of methylation were observed at least in testis at ZT13. Conclusion： Methylation of E-box CpGs does not underly the basis of rnPerl atypical rhythmic expression in testis and thymus. Sub-groups of CpGs around E-boxes 3 and 4 appears to be preferentially methylated, and might play a role in regulating mPerl transcription.
出处
《首都师范大学学报（自然科学版）》
2009年第3期33-37,共5页
Journal of Capital Normal University：Natural Science Edition
基金
国家自然科学基金(30400148
30430280
3042801040
30371574)
科技部重点项目(2006CB500701
2004BA702B02)
北京市自然科学基金项目(7031002)
北京市科技新星计划(H020821400190)
