董忠典,张宁,崔忠凯,徐文腾,陈松林

• 1:广东海洋大学水产学院
• 2:中国水产科学研究院黄海水产研究所
• 3:青岛海洋科学与技术国家实验室/海洋渔业科学与食物产出过程功能实验室

摘要(Abstract)：

【目的】研究半滑舌鳎(Cynoglossussemilaevis)gata5基因(csgata5)的序列特征及该基因在半滑舌鳎性腺分化、成熟过程中的表达模式。【方法】通过克隆获得csgata5编码区序列,利用荧光定量PCR(RT-qPCR)方法检测csgata5 mRNA在不同类型性腺、胚胎和性腺发育中的表达模式,借助原位杂交技术定位csgata5 mRNA表达的细胞类型。【结果】克隆获得csgata5 cDNA序列1 777 bp,包含1 167 bp的编码区(Coding sequence,CDS),262 bp的5′UTR和348 bp的3′UTR区,预测csgata5编码388个氨基酸残基(aa),分子质量为41.9 ku,等电点为8.90,在C端含有两个典型的GATA锌指结构域;基因组分析显示csgata5位于半滑舌鳎10号染色体上,包含7个外显子和6个内含子区。RT-qPCR分析显示csgata5 mRNA在2龄雄鱼和伪雄鱼精巢中表达高于卵巢;csgata5m RNA主要定位在卵母细胞、精原细胞和精子中。胚胎发育过程中,csgata5m RNA表达在1细胞期至囊胚期呈下降趋势,之后上升,至原肠胚期达到峰值,随后降低并维持较低水平至孵化出膜。在精巢发育过程中,csgata5m RNA表达水平呈升高趋势,在1龄达到检测的峰值;在卵巢发育过程中,csgata5m RNA表达趋势类似精巢,表达水平低于同期精巢。【结论】csgata5参与半滑舌鳎原肠胚期的组织器官分化,具有促进原始生殖细胞向雄性生殖细胞分化及雄性生殖细胞发育的作用。

关键词(KeyWords)： 半滑舌鳎;gata5;RT-qPCR;原位杂交;性腺分化

Abstract:

Keywords:

基金项目(Foundation): 黄海水产研究所科研事业单位基本科研业务费项目(20603022017003);; 山东省泰山学者攀登计划项目;; 国家自然科学基金青年科学基金项目(31402293);; 广东海洋大学博士启动项目

作者(Author): 董忠典,张宁,崔忠凯,徐文腾,陈松林

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