长白猪×蓝塘猪及其杂种基因组DNA甲基化分析  

Analysis of DNA Methylation in Landrace×Lantang Pigs and Their Hybrids Based on MSAP Marker

作　　者：肖正中[1,2] 刘小红[3] 王翀[1] 李加琪[1] 莫德林[3] 陈瑶生[3] 

机构地区：[1]华南农业大学动物科学学院,广州510642 [2]广东韶关学院农业科学系,韶关512005 [3]中山大学生命科学学院,广州510006

出　　处：《畜牧兽医学报》2011年第5期613-620,共8页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：国家自然科学基金重点项目(U0731003);广州市科技攻关重点项目(2008Z1-E121)

摘　　要：本研究旨在分析猪基因组DNA胞嘧啶甲基化模式和程度以及父母代猪与其杂种F1代之间的甲基化差异。应用甲基化敏感扩增多态性(MSAP)技术对6头长白公猪、50头蓝塘母猪及长×蓝51头杂交F1代3个群体共107个个体耳组织基因组DNA胞嘧啶甲基化模式和程度进行评估。应用筛选的20对引物扩增,总共得到1 074条带(CCGG位点),共得到415个甲基化位点(条带),其中79条带为甲基化多态性条带,平均甲基化多态性比率(p)达7.4%。结果显示,3个猪群中DNA甲基化程度高,且亲代和F1代杂种之间的甲基化水平存在差异,父母代及其杂交F1代3个群体基因组DNA甲基化总体水平分别是27.7%、27.8%和25.1%,且3个猪群中CCGG序列发生单链DNA外部胞嘧啶甲基化现象(20.8%)比单双链DNA内部胞嘧啶甲基化现象(17.9%)多。结果提示,使用MSAP技术检测猪种基因组甲基化多态性非常有效,且猪基因组甲基化多态性丰富,杂种F1代与其父母代之间的基因组甲基化水平存在差异。This experiment was conducted to analyze the extent and pattern of cytosine methylation in porcine genome and the difference of methylation between the parents and their F1 hybrids.The extent and pattern of cytosine methylation was assessed in genomes from 6 Landrace boars,50 Guangdong Lantang sows and 51 their F1 hybrids by using the technique of methylation-sensitive amplified polymorphism（MSAP）.A total of 1 074 DNA fragments including CCGG sites were amplified using 20 pairs of selective primers,in total,415 methylated bands（sites） including 79 polymorphic bands were obtained,and the average proportion of methylation polymorphism（p） was 7.4%.The results demonstrated that DNA methylation extents were high and methylation level varied among the three populations,genomic DNA methylation level of male parents,female parents and their F1 hybrids were 27.7%,27.8% and 25.1%,respectively;and the external cytosine methylation frequecy of the CCGG sequence in single strand DNA（20.8%） was higher than that in single or double strand DNA（17.9%）.The results indicate MSAP technique is efficient for detecting cytosine methylation in porcine genomes,and the methylation polymorphism in pigs is high,and the genome methylation level of F1 hybrids was different from their parents.

关 键 词：猪 DNA 甲基化 MSAP 

分 类 号：S828[农业科学—畜牧学] Q503[农业科学—畜牧兽医]