转基因克隆牛胎盘中印迹基因PEG10的DNA甲基化水平

doi:10.3724/SP.J.1005.2011.00533

遗传 ›› 2011, Vol. 33 ›› Issue (5): 533-538.doi: 10.3724/SP.J.1005.2011.00533

• 研究报告 • 上一篇

转基因克隆牛胎盘中印迹基因PEG10的DNA甲基化水平

苏建民, 许文兵, 李艳艳, 王丽君, 王勇胜, 张涌

西北农林科技大学动物医学院, 农业部动物生殖生理和胚胎工程重点实验室, 杨凌 712100

收稿日期:2010-10-14 修回日期:2010-12-09 出版日期:2011-05-20 发布日期:2011-05-25
通讯作者: 张涌 E-mail:zhy1956@263.net
基金资助:
国家转基因生物新品种培育科技重大专项(编号：2008ZX08007-004)资助

The methylation status of PEG10 in placentas of cloned transgenic calves

SU Jian-Min, XU Wen-Bing, LI Yan-Yan, WANG Li-Jun, WANG Yong-Sheng, ZHANG Yong

College of Veterinary Medicine, Northwest A&F University, Key Laboratory of Animal Reproductive Endocrinology & Embryo Bio-technology, Ministry of Agriculture, Yangling 712100, China

Received:2010-10-14 Revised:2010-12-09 Online:2011-05-20 Published:2011-05-25
Contact: ZHANG Yong E-mail:zhy1956@263.net

摘要/Abstract

摘要： 低效率的体细胞核移植技术显著制约着该技术在转基因动物生产上的广泛应用。目前认为供体细胞核不能被受体卵母细胞胞质完全的表观重编程是其效率低下的最主要原因, 而DNA甲基化是基因表观修饰的主要方式之一。为了探求转基因克隆牛的死亡是否与其胎盘中印迹基因的甲基化的重编程程度相关, 文章通过亚硫酸氢盐测序法(Bisulfite sequencing PCR, BSP)和亚硫酸氢盐联合限制性内切酶分析法(Combined bisulfite restriction analysis, COBRA), 对印迹基因PEG10在围产期死亡且存在发育缺陷的转基因克隆牛的胎盘 (死亡组)和存活的转基因克隆牛的胎盘(存活组)与正常对照牛胎盘(对照组)的DNA甲基化水平进行了详细的比较。结果发现, 与对照组相比, PEG10基因在死亡组上表现出异常的超甲基化水平, 而存活组与对照组相比无显著性差异。研究结果显示, 胎盘中印迹基因的DNA甲基化表观重编程不彻底可能是导致转基因克隆牛发育异常进而死亡的主要原因之一。

关键词: 体细胞核移植, 转基因克隆牛, PEG10, DNA甲基化

Abstract: The low efficiency of somatic cell nuclear transfer (SCNT) is a significant barrier to the production of highly valuable transgenic livestock. It is generally believed that the principal cause of the low SCNT efficiency is the aberrant nuclear epigenetic reprogramming of donor somatic cell. DNA methylation is a major epigenetic modification of the genome and plays a crucial role in nuclear reprogramming during SCNT. In order to assess whether the abnormal epigenetic modifications of the imprinted gene in placenta are correlated with the development abnormality and death of the cloned transgenic calves, the DNA methylation patterns of PEG10 were compared in the placentas from different kinds of cattle. This comparison included transgenic cloned calves died during perinatal stage and showed developmental defects (Death group), transgenic cloned calves survived and lived on healthily (Live group) and the normal reproduced calves (N group) used as the control group analyzed by Bisulfite Sequencing PCR (BSP) method and Combined Bisulfite Restriction Analysis (COBRA). Comparing to the control group, PEG10 gene in the Death group showed abnormal hyper-methylation, but was not significant different in methylation level from the Live group. It can be postulated from the results that the incomplete or abnormal DNA methylation epigenetic reprogramming of imprinting gene in placenta may be one of the main causes of the abnormal development and death of the transgenic cloned cattle.

Key words: DNA methylation, PEG10, SCNT, cloned transgenic cow

苏建民，许文兵，李艳艳，王丽君，王勇胜，张涌. 转基因克隆牛胎盘中印迹基因PEG10的DNA甲基化水平[J]. 遗传, 2011, 33(5): 533-538.

SU Jian-Min, HU Wen-Bing, LI Yan-Yan, WANG Li-Jun, WANG Yong-Qing, ZHANG Chong. The methylation status of PEG10 in placentas of cloned transgenic calves[J]. HEREDITAS, 2011, 33(5): 533-538.

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转基因克隆牛胎盘中印迹基因PEG10的DNA甲基化水平

The methylation status of PEG10 in placentas of cloned transgenic calves

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