H19和IGF2R基因在体细胞克隆猪各组织中的甲基化状态  

The methylation status of H19 and IGF2R in different tissues of somatic cell cloned pig

作　　者：吴志强[1,2,3] 谢一妮[1,2,3] 戴建军[2,3] 张廷宇[2,3] 吴彩凤[2,3] 张树山[2,3] 顾晓龙[1,2,3] 刘亮[2,3] 吴斌[1,2,3] 陈慧兰[2,3] 张德福[2,3] 马恒东[1] 

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

出　　处：《中国兽医学报》2012年第6期922-926,932,共6页Chinese Journal of Veterinary Science

基　　金：国家转基因生物新品种培育科技重大专项基金资助项目(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：为了探求新生克隆猪可能的死亡原因以及是否存在不完全的DNA甲基化重编程,本试验运用亚硫酸氢盐测序法分别检测了H19基因和IGF2R基因差异甲基化区(DMR)在新生死亡克隆猪和同期正常猪心脏、肝脏、脾脏、肺脏和肾脏中的甲基化状态。结果发现,H19基因DMR在克隆猪肺脏中表现为超甲基化,极显著高于正常猪(95.20%vs 46.80%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(4～9位、12～S17位),而在其他组织中甲基化差异不显著(P>0.05);IGF2R基因DMR在肝脏中处于超甲基化状态,显著高于正常猪(80.00%vs 39.41%P<0.05),而在肺脏中为去甲基化状态,极显著低于正常猪(14.71%vs 66.47%P<0.01),在其他组织差异不显著(P>0.05)。结果说明,在死亡克隆猪中,H19基因DMR在肺脏和IGF2R基因在肝脏与肺脏中存在不完全的DNA甲基化重编程,这可能是导致克隆动物死亡的因素之一。In order to search for the possible reasons for the death of new borned cloned pig,and whether the incom- plete DNA methylation reprogramming has occurred in SCNT animals. The DNA methylation status of H19 and IGF2R differentially methylated region （DMR） in heart,liver,spleen,lung and kidney of cloned pig and normal pig were analyzed using bisulfite sequencing analysis. The results demonstrated that the H19 DMR was hypermethylated in the lung of the dead cloned pig,and was significantly higher compared with the control （95.20% vs 46.80% P〈0.01）. Also,the tested CpGs sites from 4 to 9 and 12 to17 exhibited full methylation. The differences among others tissues were insignificant （P〉0.05） ; the IGF2R DMR showed hypermethylation in liver of the cloned pig,and was significantly higher than the control （80.00% vs 39.41% P〈0. 05）; while in lung of the dead cloned pig, the IGF2R DMR was hypomethylated, and was ignificantly lower than the control （14.71% vs 66.47 % P〈0.01）. Re- sults showed that the incomplete DNA methylation reprogramming of H19 DMR in the lung and IGF2R DMR in the lung and Hver of cloned pig might 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：S828[农业科学—畜牧学] Q78[农业科学—畜牧兽医]