实验方法 Experimental Methods

用高效液相色谱法检测鸡胚组织基因组DNA甲基化水平变化规律

  • 李世召 ,
  • 郭伟 ,
  • 杜超 ,
  • 景宇飞 ,
  • 刘艳利 ,
  • 申静 ,
  • 姚军虎 ,
  • 杨小军
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  • 西北农林科技大学动物科技学院, 杨凌 712100

收稿日期: 2014-08-24

  网络出版日期: 2015-01-10

基金资助

国家自然基金项目(31272464,31001017);教育部新世纪优秀人才(NCET-12-0476);陕西省科技新星基金(2012KJXX-18)

Analysis of Variation of Genome DNA Methylation Level of Chick Embryo Tissues with High Performance Liquid Chromatography

  • LI Shizhao ,
  • GUO Wei ,
  • DU Chao ,
  • JING Yufei ,
  • LIU Yanli ,
  • SHEN Jing ,
  • YAO Junhu ,
  • YANG Xiaojun
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  • College of Animal Science and Technology, Northwest A & F University, Yangling 712100, China

Received date: 2014-08-24

  Online published: 2015-01-10

摘要

本试验旨在优化用于基因组DNA甲基化水平检测的高效液相色谱法,并初步探索鸡胚发育过程中不同组织基因组DNA甲基化水平的变化规律。首先,试验从DNA酶解体系和高效液相色谱仪检测条件2个方面改进基因组DNA甲基化水平检测方法;然后,以孵化第8、11、14和17天的科宝500肉鸡胚胎为材料,利用改进的高效液相色谱法测定心脏、肝脏和肌肉的基因组DNA甲基化水平。结果表明:3种组织的基因组DNA甲基化水平均随胚龄增加而升高,且鸡胚发育后期(孵化第17天)肝脏的甲基化水平显著高于心脏和肌肉(P<0.05);各胚龄心脏和肌肉的基因组DNA甲基化水平均差异不显著(P>0.05)。结果提示,优化后的高效液相色谱法为快速、准确地获取组织基因组DNA甲基化水平奠定了基础;随着鸡胚发育的进行,基因组DNA甲基化水平逐渐升高,且在胚胎发育后期肝脏甲基化水平较高。

本文引用格式

李世召 , 郭伟 , 杜超 , 景宇飞 , 刘艳利 , 申静 , 姚军虎 , 杨小军 . 用高效液相色谱法检测鸡胚组织基因组DNA甲基化水平变化规律[J]. 动物营养学报, 2015 , 27(1) : 171 -177 . DOI: 10.3969/j.issn.1006-267x.2015.01.021

Abstract

This experiment was conducted to develop the high performance liquid chromatography (HPLC) method for genome DNA methylation level detection, and to explore the variation of genome DNA methylation level of chick embryo tissues preliminary. Firstly, the genome DNA methylation level detection method was improved from 2 aspects including DNA enzymatic hydrolysis system and HPLC instrument detection condition; then, the DNA methylation level of heart, liver and muscle at embryonic ages of 8, 11, 14, 17 days of Cobb 500 broiler chick embryo was detected by improved HPLC method, respectively. The results showed that the genome DNA methylation level of 3 tissues was increased with embryonic age increasing and that of liver was significantly higher than that of heart and muscle in later embryonic development (embryonic age of 17 days) (P<0.05). Heart and muscle showed no significant difference in DNA methylation level among all embryonic ages (P>0.05). It is concluded that the developed HPLC laid a good foundation for genome DNA methylation determination of tissues rapidly and accurately. Otherwise, the genome DNA methylation level rises gradually with chick embryonic age increasing, and that of liver is higher in later embryonic development.

Key words: HPLC; chick; embryo; DNA methylation

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