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中国农学通报 ›› 2014, Vol. 30 ›› Issue (35): 126-131.doi: 10.11924/j.issn.1000-6850.2014-0832

所属专题: 生物技术

• 水产 渔业 • 上一篇    下一篇

黄鳝Dmrt3基因在性腺中的甲基化差异

宋颖   

  1. 南京农业大学
  • 收稿日期:2014-03-25 修回日期:2014-03-25 接受日期:2014-05-30 出版日期:2015-03-18 发布日期:2015-03-18
  • 通讯作者: 邴旭文
  • 基金资助:
    公益性行业科研专项“鳝鱼产业技术研究与试验示范”(201003076);国家自然科学基金项目(31200918);中央级公益性科研院所基本科研业务费专项资金(中国水产科学研究院淡水渔业研究中心)项目(2013JBFM02、2011JBFA12);江苏省自然科学基金项目(BK2011184);江苏省科技支撑计划(BE2013315):江苏省水产三项工程项目(PJ2011-51)。

Dmrt3 Methylation Differences of Sex Gland in Monopterus albusGene

  • Received:2014-03-25 Revised:2014-03-25 Accepted:2014-05-30 Online:2015-03-18 Published:2015-03-18

摘要: DNA甲基化修饰是真核生物最常见的表观遗传现象之一,是一种可逆的过程,能够直接影响到基因的活性。黄鳝具有性逆转的特点,对于黄鳝的性别控制相关基因我们选择了Dmrt3基因,该基因已经证明在鱼类雌雄性腺中存在表达差异。黄鳝的Dmrt3基因编码的mRNA全长为1413bp,编码470个氨基酸。我们通过比较Dmrt3基因在雌雄性腺中甲基化状态就可以简单了解这些基因的开关状况。用甲基化敏感的限制性内切酶Hpa II酶切黄鳝的雌雄性腺,根据已知的mRNA序列设计相关引物扩增酶切产物,与未酶切的对照组进行比较,分析差异条带并验证结果,发现Dmrt3-FR2扩增序列中,精巢是去甲基化的,而卵巢是甲基化状态。所以此位点很可能与性别调控相关。本实验为黄鳝性别调控的相关研究提供一种分子学角度的方法。

关键词: 相关性, 相关性

Abstract: Monopterus albus is sex reversal, this study aims to find sequence sites directly related to sex regulation, and analyze sequence differences in male and female Monopterus albus. In this experiment, the Dmrt3 gene was taken as the research object, by comparing the gene methylation status in male and female gonads, specific sites of sex control were analyzed. Using methylation sensitive restriction enzyme Hpa II, the DNA of Monopterus albus’s gonad was cut and the enzyme digested products were amplified by each primer according to the known mRNA sequence. At the same time, the control group with no enzyme was compared with the test group, to analyze the different bands and to verify the results. The experimental results showed that Dmrt3-FR2 amplification sequences had no methylation sites in the testis, but had methylation sites in ovarian. Therefore, we speculate the site may be related to sex regulation. This experiment is a supplement to the study of sex control from the molecular angle in Monopterus albus, and proves the feasibility of using methylation to research sex control, it may provide a theoretical basis for the other researches.