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[克隆转基因犊牛胎盘组织中PEG10的甲基化状态]

[The methylation status of PEG10 in placentas of cloned transgenic calves].

作者信息

Su Jian-Min, Xu Wen-Bing, Li Yan-Yan, Wang Li-Jun, Wang Yong-Sheng, Zhang Yong

机构信息

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

出版信息

Yi Chuan. 2011 May;33(5):533-8. doi: 10.3724/sp.j.1005.2011.00533.

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 Analy-sis (COBRA). Comparing to the control group, PEG10 gene in the Death group showed abnormal hypermethylation, but was not significant different in methylation level from the Live group. It can be postulated from the results that the incom-plete 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.

摘要

体细胞核移植(SCNT)效率低下是生产高价值转基因家畜的一个重大障碍。人们普遍认为,SCNT效率低下的主要原因是供体体细胞的核表观遗传重编程异常。DNA甲基化是基因组的一种主要表观遗传修饰,在SCNT过程中的核重编程中起着关键作用。为了评估胎盘中印记基因的异常表观遗传修饰是否与克隆转基因犊牛的发育异常和死亡相关,采用亚硫酸氢盐测序PCR(BSP)法和联合亚硫酸氢盐限制性分析(COBRA),比较了不同种类牛胎盘组织中PEG10基因的DNA甲基化模式。研究对象包括围产期死亡且有发育缺陷的转基因克隆犊牛(死亡组)、存活且健康生长的转基因克隆犊牛(存活组)以及作为对照组的正常繁殖犊牛(N组)。与对照组相比,死亡组的PEG10基因呈现异常的高甲基化,但与存活组在甲基化水平上无显著差异。从结果可以推测,胎盘组织中印记基因的DNA甲基化表观遗传重编程不完全或异常可能是转基因克隆牛发育异常和死亡的主要原因之一。

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