牛体细胞核移植中Meg8基因DNA甲基化及印迹状态分析

doi:10.11843/j.issn.0366-6964.2014.09.006

畜牧兽医学报 ›› 2014, Vol. 45 ›› Issue (9): 1417-1423.doi: 10.11843/j.issn.0366-6964.2014.09.006

牛体细胞核移植中Meg8基因DNA甲基化及印迹状态分析

赵姝君¹，胡嘉祺¹，张明月¹，李冬杰²，戴蕴平³，李宁³，李世杰^1*

(1．河北农业大学生命科学学院，保定 071001； 2．河北科技大学生物科学与工程学院，石家庄 050018；3．中国农业大学农业生物技术国家重点实验室，北京 100193)

收稿日期:2014-03-11 出版日期:2014-09-23 发布日期:2014-09-23
通讯作者: 李世杰，教授，博士生导师，Tel：0312-7528251，E-mail：lishijie20005@163.com
作者简介:赵姝君(1988-)，女，河北石家庄人，硕士生，主要从事动物分子遗传方面的研究，Tel：0312-7528270，E-mail:niansuiusi0718 @163.com
基金资助:
国家自然科学基金项目(31372312)；河北省自然基金项目（C2011204001）

DNA Methylation and Imprinting Status of Meg8 in Somatic Cell Nuclear Transfer Bovines

ZHAO Shu-jun¹，HU Jia-qi¹，ZHANG Ming-yue¹，LI Dong-jie²，DAI Yun-ping³，LI Ning³ ，LI Shi-jie^1*

(1.College of Life Science，Agricultural University of Hebei，Baoding 071001，China;2.College of Bioscience & Bioengineering，Hebei University of Science and Technology，Shijiazhuang 050018，China;3.State Key Laboratory for Agrobiotechnology,China Agricultural University， Beijing 100193,China)

Received:2014-03-11 Online:2014-09-23 Published:2014-09-23

摘要/Abstract

摘要：

为了确定Meg8基因内部CpG岛的甲基化在调控Meg8基因印记中的可能作用，本研究应用亚硫酸盐测序法分析Meg8基因内含子3上的17个CpGs位点等位基因特异的DNA甲基化状态。结果发现，2条亲本链的甲基化程度都比较高（>80%），但T链甲基化水平显著高于C链（P<0.05），并且C链只有1种甲基化模式。分析Meg8基因在出生后48 h死亡的体细胞核移植牛肺中的甲基化和印迹状态，发现Meg8基因甲基化水平在核移植牛和自然繁殖牛中都比较高，但核移植牛的甲基化程度显著高于正常繁殖牛（P<0.05），并且只有1种完全甲基化的模式，而Meg8基因在核移植个体中的印记表达没有紊乱，仍表现为单等位基因表达，初步推测Meg8内含子3CpGs岛的甲基化可能没有参与调控Meg8基因的印记表达。

Abstract:

In order to determine the possible roles of Meg8 gene-internal CpG island in the regulation of the gene imprinting expression，the allele-specific DNA methylation status of 17 CpGs in intron3 of Meg8 in lung tissues was analyzed using bisulfite sequencing.The results demonstrated that the DNA methylation of both the parental strands were high （>80%），but T-strand showed significantly higher DNA methylation than that of C-strand（P<0.05） and the C-strand had only one kind of methylation model.Then the DNA methylation and imprinting status were analyzed in lung in somatic cell nuclear transfer(SCNT) bovine which died within postnatal 48 h.The results showed that the SCNT bovines exhibited significantly higher DNA methylation than that of naturally breed bovines（P<0.05）.All of 17 CpGs showed complete methylation pattern in SCNT bovine.Imprinting analysis showed that Meg8 was mono-allelic expression in tissues of SCNT bovines.The DNA methylation of Meg8 intron3 CpGs island may not play a role in regulating of Meg8 imprinting expression.

中图分类号:

赵姝君，胡嘉祺，张明月，李冬杰，戴蕴平，李宁，李世杰. 牛体细胞核移植中Meg8基因DNA甲基化及印迹状态分析[J]. 畜牧兽医学报, 2014, 45(9): 1417-1423.

ZHAO Shu-jun，HU Jia-qi，ZHANG Ming-yue，LI Dong-jie，DAI Yun-ping，LI Ning,LI Shi-jie. DNA Methylation and Imprinting Status of Meg8 in Somatic Cell Nuclear Transfer Bovines[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2014, 45(9): 1417-1423.

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牛体细胞核移植中Meg8基因DNA甲基化及印迹状态分析

DNA Methylation and Imprinting Status of Meg8 in Somatic Cell Nuclear Transfer Bovines

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