绵羊成纤维细胞及精子Sox2基因启动子甲基化状态检测  

Detection of methylation status of Sox2 gene promoter in sheep fibroblasts and sperms

作　　者：陈胜男 宋红卫[1] 李松美 朴善花[1] 王春生[1] CHEN Shengnan;SONG Hongwei;LI Songmei;PIAO Shanhua;WANG Chunsheng(School of Life Sciences,Northeast Forestry University,Harbin 150040,China)

机构地区：[1]东北林业大学生命科学学院,哈尔滨150040

出　　处：《黑龙江畜牧兽医》2020年第1期22-26,共5页Heilongjiang Animal Science And veterinary Medicine

基　　金：中央高校基本科研业务费专项(2572016CA10);黑龙江省自然科学基金项目(C2016012)。

摘　　要：为了探明绵羊成纤维细胞及精子中Sox2基因启动子甲基化状态的差异,试验对绵羊Sox2基因启动子进行序列查询和甲基化岛分析,提取绵羊成纤维细胞和精子基因组,用重亚硫酸盐处理,进行甲基化特异性PCR、连接和酶切鉴定,最终分别随机选取10个测序结果进行甲基化状态分析,比较二者的甲基化状态。结果表明:在精子的350个甲基化位点中,有194个位点处于非甲基化状态,156个位点处于甲基化状态,甲基化率为44.6%;在成纤维细胞的350个甲基化位点中,93个位点处于非甲基化状态,257个位点处于甲基化状态,甲基化率为73.4%;χ~2检验表明二者的甲基化状态差异极显著(P<0.01)。说明绵羊精子和成纤维细胞的Sox2基因启动子区域甲基化状态存在较大差异。To investigate the differences in methylation status of Sox2 gene promoters in fibroblasts and sperms,the query of the Sox2 promoter sequence of sheep and methylation island analysis were performed.The genome was extracted from sheep fibroblasts and sperm,and treated with bisulfite.Methylation-specific PCR,ligation,and restriction enzyme digestion were conducted.Finally,we selected randomly 10 sequencing results,performed methylation status analysis,and compared methylation state.The results showed that 194 of the 350 sites in the sperms were unmethylated;156 were methylated;and the methylation rate was 44.6%.In 350 methylation sites,93 sites were unmethylated;257 sites were methylated;and the methylation rate was 73.4%.The chi-square test showed that the difference was extremely significant(P<0.01).These data indicate that the methylation status of promoter region of Sox2,a pluripotency gene,is quite different in sheep fibroblasts and sperms.

关 键 词：绵羊精子 绵羊成纤维细胞 Sox2基因 启动子 DNA甲基化 

分 类 号：S826[农业科学—畜牧学] Q78[农业科学—畜牧兽医]