克隆猪印迹基因IGF2/H19印迹控制区的甲基化状态

The Methylation Status of IGF2/H19 Imprinting Control Region in Cloned Pig

为了探求核移植过程中DNA甲基化重编程是否充分,运用亚硫酸氢盐测序法分别检测新生死亡克隆猪和同期正常猪心脏、肝脏、脾脏、肺脏和肾脏组织中IGF2/H19基因印迹控制区(DMR1、DMR2、DMR3)的甲基化状态。结果发现,DMR1、DMR3在克隆猪和正常猪各组织中的甲基化水平不同,但差异不显著(P>0.05)。DMR2在克隆猪肺脏组织表现为超甲基化,极显著高于正常猪(P<0.01),且10个测序克隆中存在2处连续的全甲基化CpG位点(分别为4-9位和12-17位),而在其它组织中甲基化差异不显著(P>0.05)。说明DMR2在克隆猪肺脏组织可能存在DNA甲基化重编程紊乱,这也可能是导致该克隆猪死亡的因素之一。

In order to make sure whether the DNA methylation reprogramming is efficient in SCNT animals,we analyzed the DNA methylation status of IGF2/H19 imprinting control region（DMR1,DMR2,DMR3） in heart,liver,spleen,lung and kidney of cloned pig using bisulfite sequencing analysis.The results demonstrated that the methylation level of DMR1 and DMR3 in dead cloned pig tissues were different from that of normal pig,but there was no significant differences among them（P〉0.05）;The DMR2 showed hypermethylation in the lung of the dead cloned pig,and was significantly higher than that in the control（P〉0.01）.Also,the tested CpGs sites from 4 to 9 and 12 to 17 exhibited full methylation.There was no significant differences among other tissues in the DMR2（P〉0.05）.Results showed that the abnormal DNA methylation proflies of DMR2 may occurred in the lung of cloned pig,which may be one of the factors for the death of cloned animals.