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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

DOI: 10.3969/j.issn.1006-267x.2015.01.021

Keywords: 高效液相色谱,肉鸡,胚胎,DNA甲基化

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Abstract:

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。

References

[1]  ROSENQUIST T H,FINNELL R H.Genes,folate and homocysteine in embryonic development[J].Proceedings of the Nutrition Society,2001,60(1):53-61.
[2]  HOLLINGWWORTH J W,MARUOKA S,BOON K,et al.In utero supplementation with methyl donors enhances allergic airway disease in mice[J].The Journal of Clinical Investigation,2008,118(10):3462-3469.
[3]  SKINNER M K,HAQUE C G B M,NILSSON E,et al.Environmentally induced transgenerational epigenetic reprogramming of primordial germ cells and the subsequent germ line[J].PLoS One,2013,8(7):e66318.
[4]  HERMAN J G,GRAFF J R,MY?H?NEN S,et al.Methylation-specific PCR:a novel PCR assay for methylation status of CpG islands[J].Proceedings of the National Academy of Sciences of the United States of America,1996,93(18),9821-9826.
[5]  GONZALGO M L,LIANG G N,SPRUCK Ⅲ C H,et al.Identification and characterization of differentially methylated regions of genomic DNA by methylation-sensitive arbitrarily primed PCR[J].Cancer Research,1997,57(4):594-599.
[6]  YANG A S,ESTECIO M R H,DOSHI K,et al.A simple method for estimating global DNA methylation using bisulfite PCR of repetitive DNA elements[J].Nucleic Acids Research,2004,32(3):e38.
[7]  彭思远,张洁,田美平,等.液相色谱-串联质谱法测定生物样本全基因组DNA甲基化[J].分析化学,2012,40(8):1201-1206.
[8]  尹慧,黄代新,翟仙敦,等.HPLC法检测人外周血5-mC含量及其与年龄相关性研究[J].中国法医学杂志,2007,22(1):8-11.
[9]  CRAIN P F.Preparation and enzymatic hydrolysis of DNA and RNA for mass spectrometry[J].Methods in Enzymology,1990,193:782-790.
[10]  LI Q H,LI N,HU X X,et al.Genome-wide mapping of DNA methylation in chicken[J].PLoS One,2011,6(5):e19428
[11]  班谦.家鸡胚胎早期发育过程中DNA甲基化的MSAP分析[D].硕士学位论文.石河子:石河子大学,2009.
[12]  唐韶青,张沅,徐青,等.不同动物部分组织基因组甲基化程度的差异分析[J].农业生物技术学报,2006,14(4):507-511.

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