斜带石斑鱼卷轴受体3基因的分离及表达分析研究

发布时间：2018-03-16 01:26

本文选题：斜带石斑鱼　切入点：卷轴受体基因　出处：《中国海洋大学学报(自然科学版)》2017年01期 　论文类型：期刊论文

【摘要】：利用cDNA末端快速扩增技术(Rapid amplification of cDNA ends,RACE)克隆鉴定出斜带石斑鱼(Epinephelus coioides)卷轴受体3(Frizzled 3,gFZD3)基因的cDNA序列全长,使用ClustalX(1.81)和MEGA4.0软件分别进行了序列同源比对和系统进化树分析;利用RT-PCR检测gFZD3在成鱼不同组织中的表达和gFZD3在胚胎发育及卵巢发育过程中的表达模式。研究显示:gFZD3cDNA序列全长为2 278bp,其中开放读码框为2 079bp,编码692个氨基酸;序列同源性分析表明,gFZD3的氨基酸序列具有FZD家族保守的功能结构域;脊椎动物的FZD3系统进化树分析显示,斜带石斑鱼与大黄鱼(Larimichthys crocea)的亲缘关系最近;在成鱼的组织分布中,只在脑区和性腺中检测到gFZD3表达信号,其中在卵巢中表达量最高;在胚胎发育过程中,在未受精卵和受精卵中gFZD3的表达量最高,随着胚胎的发育呈逐渐下降趋势;在卵巢发育过程中,gFZD3的表达在早期卵巢中相对较低,在随后的III和IV期卵巢中较高。研究结果表明,FZD3序列在脊椎动物中具有较高的保守性;斜带石斑鱼gFZD3的表达具有明显的组织特异性,并在胚胎和卵巢发育中,可能主要作用于胚胎发育的早期和卵巢发育的后期。本研究为进一步研究Wnt信号通路在斜带石斑鱼生殖调控中的作用提供了参考。
[Abstract]:Rapid amplification of cDNA endsrase (RACEE) was used to clone and identify the full length of the cDNA gene of Epinephelus coioides-3 (Frizzled 3gFZD3) gene. The sequence homology alignment and phylogenetic tree analysis were performed by ClustalXS1.81) and MEGA4.0 software, respectively. RT-PCR was used to detect the expression of gFZD3 in different tissues of adult fish and the expression pattern of gFZD3 in embryonic development and ovarian development. The results showed that the full length of the cDNA sequence was 2278bp. the open reading frame was 2079bp, encoding 692 amino acids. Sequence homology analysis showed that the amino acid sequence of GFZD3 had the conserved functional domain of FZD family, and the FZD3 phylogenetic tree analysis of vertebrates showed that there was the closest phylogenetic relationship between the oblique grouper and Larimichthys Crocea, and in the tissue distribution of adult fish, The expression of gFZD3 was detected only in the brain and gonad, the highest in ovary, the highest in unfertilized egg and fertilized egg during embryonic development, and decreased gradually with the development of embryo. The expression of gFZD3 was relatively low in the early stage of ovarian development and higher in the subsequent stages of III and IV. The results showed that the FZD3 sequence was highly conserved in vertebrates. The expression of gFZD3 in Epinephelus obliquus has obvious tissue specificity and is found in embryonic and ovarian development. This study may play an important role in the early stage of embryonic development and the late stage of ovarian development. This study provides a reference for further study on the role of Wnt signaling pathway in the reproductive regulation of Epinephelus oblique.
【作者单位】：广东海洋大学水产学院海洋生态与养殖环境湛江市重点实验室;三亚市海洋与渔业监测中心;
【基金】：广东省海洋渔业科技推广专项(A201408A06) 海南省科技合作专项(KJHZ2015-08)资助~~
【分类号】：S917.4

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