ES小鼠和ES细胞中H19基因甲基化状态

为探讨印迹基因H19的甲基化状态与ES小鼠胚胎发育之间的关系, 以遗传背景相同的正常成年对照小鼠、22只成年ES小鼠和8只新生死亡的ES小鼠以及不同传代次数的ES细胞为实验材料, 利用甲基化敏感性限制性内切酶-PCR技术分别检测了其印迹基因H19的5′非翻译区两个位点的甲基化状态。结果表明, 发育至成年的ES小鼠印迹基因H19所检测位点的甲基化状态与正常成年对照小鼠之间没有差异, 而新生死亡的ES小鼠印迹基因H19所检测位点的甲基化状态与成年ES小鼠以及正常成年对照小鼠相比则存在明显差异。推测ES细胞中印迹基因H19所检测位点的甲基化状态与成年ES小鼠以及正常成年对照小鼠之间可能存在 差异。

Methylation of H19 gene in ES mice and embryonic stem cells

To investigate the relationship between the methylation status of imprinted gene H19 and the development of mice derived completely from Embryonic stem cells (ES) by tetraploid embryo complementation. The methylation status of two loci at 5'UTR region in imprinted gene H19 in normal adult control mice, 22 adult ES mice, 8 newborn dead ES mice and embryonic stem (ES) cells with different passage number were detected by using of methylation-sensitive restriction endonuclease-PCR technique. The results indicated that the methylation status of the imprinted gene H19 in adult ES mice were identical to that of adult control mice. However, some significant differences in the methylation status of the imprinted gene H19 were found among newborn dead ES mice, adult ES mice and normal adult control mice. Furthermore, the methylation status of the imprinted gene H19 in ES cells were probably different from that of adult ES mice and normal adult control mice.