鹿茸干细胞基因组DNA甲基化检测方法的比较
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- 英文论文题名：Comparison the methods using in detect Genome-wide DNA methylation of Antler stem cell
- 论文作者：杨春 ; 褚文辉 ; 路晓 ; 李春义
- 英文论文作者：Yang Chun ; Wenhui Chu ; Lu Xiao ; Chunyi Li ; State Key Laboratory for Molecular Biology of Special Economic Animals ; Institute of Special Animal and Sciences ; Chinese Academy of Agricultural Sciences
- 年：2016
- 作者机构：中国农业科学院特产研究所吉林省特种动物分子生物学省部共建国家重点实验室;
- 论文关键词：甲基化敏感扩增多态性 ; 荧光标记甲基化敏感扩增多态性 ; 甲基化 ; 鹿茸干细胞
- 英文论文关键词：MSAP ; F-MSAP ; DNA methylation ; antler stem cell
- 会议召开时间：2016-06-01
- 会议录名称：“十二五” 鹿科学研究进展
- 语种：中文
- 分类号：S825
- 学会代码：ZXMY
- 学会名称：中国畜牧业协会
- 页数：9
- 文件大小：996k
- 原文格式：O
摘要
本研究旨在应用甲基化敏感性扩增多态性(MSAP)和荧光标记的甲基化敏感性扩增多态性(F-MSAP)两种技术对鹿茸组织进行全基因组DNA甲基化检测,通过实验结果对两种方法进行比较,为今后鹿茸干细胞基因组DNA甲基化分析奠定基础。甲基化敏感性扩增多态性(MSAP)技术是一种从全基因组水平检测DNA甲基化状态的方法,是一种经过改进的扩增片段长度多态性(AFLP)技术。荧光标记的甲基化敏感性扩增多态性(F-MSAP)技术也是一种经过改进的AFLP方法,将选择性扩增引物进行了荧光标记,这样就可以利用测序仪自动完成实验。结果表明,F-MSAP方法具有安全、高效、高通量和自动化等特点。F-MSAP方法更适合于检测全基因组胞嘧啶甲基化。
In the study,we used The methylation- sensitive amplification polymorphism(MSAP) and fluorescence- labeled methylation- sensitive amplification polymorphism(F —MSAP) methods to analysis of DNA methylation in antler stem cells,and according to the results to compare the MSAP and F- MSAP methods.The MSAP is a modified AFLP(amplified fragment length polymorphism) technique to investigate cytosine methylation in genomes.F-MSAP is a new modification of MSAP technique,in which selective amplification is performed with fluorescently labeled and the selective products eoulde detected by DNA sequencer.In the study,we used MSAP and F- MSAP methods to analysis of DNA methylation in antler stem cells,and according to the results to compare the MSAP and F- MSAP methods.The results shown that compared to MSAP method,the F- MSAP method has the main advantages of safety,time efficiency,high sensitivity and automation.The F- MSAP method was more suitable for detection of cytosine methylation of the whole genome.
In the study,we used The methylation- sensitive amplification polymorphism(MSAP) and fluorescence- labeled methylation- sensitive amplification polymorphism(F —MSAP) methods to analysis of DNA methylation in antler stem cells,and according to the results to compare the MSAP and F- MSAP methods.The MSAP is a modified AFLP(amplified fragment length polymorphism) technique to investigate cytosine methylation in genomes.F-MSAP is a new modification of MSAP technique,in which selective amplification is performed with fluorescently labeled and the selective products eoulde detected by DNA sequencer.In the study,we used MSAP and F- MSAP methods to analysis of DNA methylation in antler stem cells,and according to the results to compare the MSAP and F- MSAP methods.The results shown that compared to MSAP method,the F- MSAP method has the main advantages of safety,time efficiency,high sensitivity and automation.The F- MSAP method was more suitable for detection of cytosine methylation of the whole genome.
引文
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[13]朱红菊,刘文革,赵胜杰,路绪强,何楠,豆峻岭,高磊。NaCI胁迫下二倍体和同源四倍体西瓜幼苗DNA甲基化差异分析[J].中国农业科学,2014,47(20):4045-4055
[14]Xu Q,Zhang Y,Sun D,Wang Y,Yu Y.Analysis on DNA methylation ofvarious tissues in chicken[J].Anim Biotechno,2007:231-241.
[15]Yonghong Zhang,Jiang Guo,Yan Gao,Shuling Niu,Chun Yang,Chunyan Bai,Xiaozhong Yu,Zhihui Zhao.Genome-wide methylation changes are associated with muscle fiber density and drip loss in male three-yellow chickens[J].MolBiol Rep,2014;3 214-3216.
[16]Yang C,Zhang MJ,Niu WP,Yang RJ,Zhang YH.Analysis of DNA Methylation in Various Swine Tissues[J].Plos One,2011,6(1):1-9.
[17]Lu YL.,Rong TZ,Cao MJ.Analysis of DNA methylationin different maize tissues[J].J Genet Genomics,2008,35(1):41-48.
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[2]Yakushiji N,Suzuki M,Satoh A,Sagai T,Shiroishi T,et al.Correlation between Shh expression and DNA methylation status of the limb-specific Shh enhancer region during limb regeneration in amphibians[J].Developmental Biology,2007,312;171-182.
[3]Iskandar BJ,Rizk E,Meier B,Hariharan N,Bottiglieri T,et al.Folate regulation of axonal regeneration in the rodent central nervous system through DNA methylation[J].Journal of Clinical Investigation,2010,120:1603-1616.
[4]Hirose K,Shimoda N,Kikuchi Y.Transient reduction of 5-methylcytosine and 5-hydroxymethylcytosine is associated with active DNA demethylation during regeneration of zebrafish fin[J].Epigenetics,2013,8:899-906.
[5]Powell C,Elsaeidi F,Goldman D.Injury-dependent Muller glia and ganglion cell reprogramming during tissue regeneration requires Apobec2a and Apobec2b[J].J Neurosci 2012,32;1 096-1 109.
[6]Tyaqi SC,Joshua IG.Exercise and nutrition in myocardial matrix metabolism,remodeling,regeneration,epigenetics,microcirculation,and muscle[J].Can J Physiol Pharmacol,2014,92:521.
[7]Qi Shen,Hongchuan Jin,Xian Wang.Epidermal stem cells and their epigenetic regulation[J].Int.J.Mol.Sci,2013,14:17861-17880.
[8]Stocum DL.Regenerative biology and medicine,2012,Academic Press.
[9]Li CY,Mackintosh CG,Martin SK,Clark DE.Identification of key tissue type for antler regeneration through pedicle periosteum deletion[J].Cell and Tissue Research,2007,328:65-75.
[10]Li CY,Yang FH,Sheppard A.Adult Stem Cells and Mammalian Epimorphic Regeneration-Insights from Studying Annual Renewal of Deer Antlers[J].Current Stem Cell Research&Therapy,2009,4:237-251.
[11]张梅,陈佳林,周晓穗,梁士可,李广宏,王方海.MSAP在水稻害虫白背飞虱中的应用研究[J].中山大学学报,2015,54(1):98-102.
[12]李金龙,唐韶青,赵萌,王海潮,徐青。北京油鸡肌肉和卵巢组织基因组DNA甲基化状态检测与分析[J].畜牧兽医学报,2014,45(11):1784-1792.
[13]朱红菊,刘文革,赵胜杰,路绪强,何楠,豆峻岭,高磊。NaCI胁迫下二倍体和同源四倍体西瓜幼苗DNA甲基化差异分析[J].中国农业科学,2014,47(20):4045-4055
[14]Xu Q,Zhang Y,Sun D,Wang Y,Yu Y.Analysis on DNA methylation ofvarious tissues in chicken[J].Anim Biotechno,2007:231-241.
[15]Yonghong Zhang,Jiang Guo,Yan Gao,Shuling Niu,Chun Yang,Chunyan Bai,Xiaozhong Yu,Zhihui Zhao.Genome-wide methylation changes are associated with muscle fiber density and drip loss in male three-yellow chickens[J].MolBiol Rep,2014;3 214-3216.
[16]Yang C,Zhang MJ,Niu WP,Yang RJ,Zhang YH.Analysis of DNA Methylation in Various Swine Tissues[J].Plos One,2011,6(1):1-9.
[17]Lu YL.,Rong TZ,Cao MJ.Analysis of DNA methylationin different maize tissues[J].J Genet Genomics,2008,35(1):41-48.
[18]Vanyushin BF.Enzymatic DNA methylation is an epigenetic control forgenetic functions of the cell[J].Biochemistry(Mosc),2005,70:488-499.
[19]Huang J,Sun M.A modified AFLP with fluorescence-labelled primersand automated DNA sequencer detection for efficient fingerprinting analysis inplants[J].Biotechnol Tech,1999,13;277-278.