应用MSAP技术研究扇贝全基因组DNA甲基化水平  

Application of MSAP Technique for Investigation of Genome-Wide DNA Methylation Level in Scallops

作　　者：吕佳[1] 侯睿[1] 李宁[1] 王宸[1] 张玲玲[1] 胡晓丽[1] 王师[1] 包振民[1] 

机构地区：[1]中国海洋大学海洋生物遗传育种教育部重点实验室,山东青岛266003

出　　处：《中国海洋大学学报（自然科学版）》2013年第10期48-53,共6页Periodical of Ocean University of China

基　　金：中央高校基本科研业务费专项;国家科技支撑计划项目(2011BAD13B01);山东省自然科学基金项目(ZR2009DM019);中国海洋大学水生生物种质资源标准化及资源共享项目资助

摘　　要：DNA甲基化作为真核生物基因组重要的表观遗传学修饰,对生物体基因的表达有重要的调控作用。为获得扇贝基因组DNA甲基化修饰水平及模式等表观遗传学信息,以栉孔扇贝(Chlamys farreri)、海湾扇贝(Argopecten irradians)、虾夷扇贝(Mizuhopecten yessoensis)以及本课题组培育的"海大金贝"为材料,建立了甲基化敏感扩增多态性方法(Methylation-sensitive amplified polymorphism,MSAP)的反应体系,利用该方法对其基因组DNA CCGG区域的甲基化水平进行分析。结果表明,本研究筛选得到的引物组合可用于贝类DNA甲基化的研究,在栉孔扇贝、海湾扇贝、普通虾夷扇贝和"海大金贝"的甲基化比例分别为32.08%、25.99%、32.88%和34.97%。几种扇贝基因组CCGG序列中,胞嘧啶的全甲基化率要高于半甲基化率,推测扇贝基因组中主要的甲基化模式是CpG型。通过对"海大金贝"和普通虾夷扇贝闭壳肌甲基化谱进行比较,筛查得到46个差异位点,这些位点可能参与"海大金贝"闭壳肌积累类胡萝卜素的调控。As an important epigenetic modification factor in eukaryotic genomes, DNA methylation plays a vital role in regulation of gene expression. In order to investigate the genome-wide DNA methylation level in molluscs, methylation-sensitive amplified polymorphism （MSAP） technique was applied to analyzing the methylation pattern at CCGG sites in three speices of scallops （Chlamys farreri, Argopecten irradi- ans, Mizuhopecten yessoensis） and ＂Haida golden scallop＂, a variety of M. yessoensis newly developed by our group. The results provided suitable MSAP primer combinations for further epigenetic studies on mol- luscs DNA methylation. The methylation level of C. farreri, A. irradians, M. yesoensis and ＂Haida golden scallop＂ wase 32.08%, 25. 990%, 32.88%, and 34. 97%, respectively. The fully-methylated sites were more than the hemi-methylated sites in these scallop species. Our data indicated that the major form of DNA methylation in scallop was CpG methylation. The comparison between ＂Haida golden scallop＂ and the common ones indicated that 46 sites were different in methylation patterns, which may be involved in the regulation of the carotenoid accumulation.

关 键 词：扇贝 DNA甲基化 甲基化敏感扩增多态性 

分 类 号：S917.4[农业科学—水产科学]