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

作　　者：吴志强[1,2,3] 谢一妮[1,2,3] 张廷宇[1,2] 戴建军[1,2] 吴彩凤[1,2] 马恒东[3] 张德福[1,2] 

机构地区：[1]四川农业大学生命科学与理学院,四川雅安625014 [2]上海农业科学院畜牧兽医研究所,上海201106 [3]上海农业遗传育种重点实验室动物遗传工程研究室,上海201106

出　　处：《中国畜牧兽医》2011年第11期125-131,共7页China Animal Husbandry & Veterinary Medicine

基　　金：国家转基因生物新品种培育科技重大专项(2008ZX08006-005;2009ZX08006-014B);上海市科技兴农攻关项目(沪农科攻字2005-3-5);上海市科技兴农推广项目(沪农科推字2007-3-7);上海市科委科技成果转化项目(103919N1800)

摘　　要：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.

关 键 词：差异甲基化区 DNA甲基化 体细胞克隆 表观重编程 

分 类 号：Q74[生物学—分子生物学]