DNA甲基化酶抑制剂5-Aza-CdR对多能基因及甲基化相关基因在徒手克隆胚胎中表达模式的影响

doi:10.16431/j.cnki.1671-7236.2016.09.004

《中国畜牧兽医》 ›› 2016, Vol. 43 ›› Issue (9): 2240-2248.doi: 10.16431/j.cnki.1671-7236.2016.09.004

DNA甲基化酶抑制剂5-Aza-CdR对多能基因及甲基化相关基因在徒手克隆胚胎中表达模式的影响

陆杏蓉^1,2, 李志鹏¹, 孙俊铭¹, 吕玲燕³, 许钟峯¹, 刘庆友¹, 石德顺¹, 崔奎青¹

1. 广西大学亚热带生物资源保护利用国家重点实验室, 南宁 530005;
2. 中国农业科学院广西水牛研究所, 广西水牛遗传繁育重点实验室, 南宁 530001;
3. 广西壮族自治区畜牧研究所, 南宁 530001

收稿日期:2016-02-15 出版日期:2016-09-20 发布日期:2016-09-20
通讯作者: 石德顺, 崔奎青 E-mail:ardsshi@gxu.edu.cn;kqcui@126.com
作者简介:陆杏蓉(1987-),女,广西百色人,硕士,研究方向:动物胚胎技术,E-mail:luxingrong074@163.com;李志鹏(1989-),男,河北邯郸人,博士生,研究方向:动物繁殖生物技术,E-mail:842209897@qq.com
基金资助:
国家自然基金（31260552）；国家高技术研究发展计划（863）项目（2011AA100607）

Effect of DNA Methylation Inhibitor 5-Aza-CdR on Patterns of Pluripotent-related Genes and Methylation Genes in Handmade Cloning Embryo

LU Xing-rong^1,2, LI Zhi-Peng¹, SUN Jun-ming¹, LV Ling-yan³, XU Zhong-feng¹, LIU Qing-you¹, SHI De-shun¹, CUI Kui-qing¹

1. State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, Guangxi University, Nanning 530005, China;
2. Guangxi Key Laboratory of Buffalo Genetics, Breeding and Reproduction, Guangxi Buffalo Research Institute, Chinese Academy of Agricultural Sciences, Nanning 530001, China;
3. Guangxi Zhuang Autonomous Region Animal Husbandry Institute, Nanning 530001, China

Received:2016-02-15 Online:2016-09-20 Published:2016-09-20

摘要/Abstract

摘要：

本试验通过实时荧光定量PCR方法对多能基因Oct4和Nanog及DNA甲基化相关基因Dnmt1和Tets在徒手克隆（handmade cloning，HMC）胚胎中的表达模式进行初步研究，并探讨5-Aza-CdR处理重构胚对这些基因表达模式的影响。结果显示，Oct4、Nanog和Tet3的表达在2细胞时期达到顶峰，Dnmt1和Tet2基因的表达随HMC胚胎发育而下降，而Tet1基因随HMC胚胎发育表达上升。使用5-Aza-CdR处理重构胚没有改变Oct4、Tet1和Tet3基因的表达模式，使Nanog基因在胚胎发育初期表达增加，Dnmt1和Tet2基因在胚胎发育初期表达降低。研究初步确立了Oct4、Nanog、Dnmt1和Tets基因在HMC胚胎的表达模式，5-Aza-CdR对重构胚的处理可对HMC胚胎的甲基化模式产生影响。

关键词: 徒手克隆; 5-Aza-CdR; 基因; 表达模式

Abstract:

The expression pattern of pluripotent gene Oct4 and Nanog,and methylation related genes Dnmt1 and Tets of handmade cloning (HMC) embryos were studied by Real-time quantitative PCR assay.The effect of genes expression pattern by 5-Aza-CdR on HMC reconstructed embryos was also explored. The results showed that the expression of Oct4,Nanog and Tet3 genes reached peak on 2-cell stage,the expression of Dnmt1 and Tet2 genes declined with the embryo development,while the expression of Tet1 gene increased.The use of 5-Aza-CdR didn't change the expression pattern of Oct4,Tet1 and Tet3 genes,but increased the expression of Nanog gene at the beginning of embryo development,while decreased the expression of Dnmt1 and Tet2 genes.The expression pattern of Oct4,Nanog,Dnmt1 and Tets genes in the development of HMC embryo was established,the use of 5-Aza-CdR could influence the methylation process of HMC embryo.

Key words: HMC; 5-Aza-CdR; gene; expression pattern

中图分类号:

S814.6

陆杏蓉, 李志鹏, 孙俊铭, 吕玲燕, 许钟峯, 刘庆友, 石德顺, 崔奎青. DNA甲基化酶抑制剂5-Aza-CdR对多能基因及甲基化相关基因在徒手克隆胚胎中表达模式的影响[J]. 《中国畜牧兽医》, 2016, 43(9): 2240-2248.

LU Xing-rong, LI Zhi-Peng, SUN Jun-ming, LV Ling-yan, XU Zhong-feng, LIU Qing-you, SHI De-shun, CUI Kui-qing. Effect of DNA Methylation Inhibitor 5-Aza-CdR on Patterns of Pluripotent-related Genes and Methylation Genes in Handmade Cloning Embryo[J]. , 2016, 43(9): 2240-2248.

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DNA甲基化酶抑制剂5-Aza-CdR对多能基因及甲基化相关基因在徒手克隆胚胎中表达模式的影响

Effect of DNA Methylation Inhibitor 5-Aza-CdR on Patterns of Pluripotent-related Genes and Methylation Genes in Handmade Cloning Embryo

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