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王晓梅,朱国利,唐文乔.2017.长江刀鲚mor-4k13基因的分离鉴定及表达分析.动物学杂志,52(1):64-74.
长江刀鲚mor-4k13基因的分离鉴定及表达分析
Isolation, Identification and Expression of mor-4k13 Gene in Japanese Grenadier Anchovy (Coilia nasus) from the Yangtze River
投稿时间:2016-03-21  修订日期:2016-12-30
DOI:10.13859/j.cjz.201701008
中文关键词:  刀鲚  嗅觉受体基因  序列分析  组织表达  生态型
英文关键词:Japanese Grenadier Anchovy, Coilia nasus  Olfactory receptor gene  Sequence analysis  Tissues expression  Ecotype
基金项目:公益性行业(农业)科研专项(No. 201203065),国家自然科学基金项目(No. 31172407,31472280),高等学校博士学科点科研专项(No. 20123104110006),上海高校水产学高峰学科;
作者单位E-mail
王晓梅 上海海洋大学 xmw0212@163.com 
朱国利 上海海洋大学  
唐文乔 上海海洋大学 wqtang@shou.edu.cn 
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中文摘要:
      分布于长江的刀鲚(Coilia nasus)具有洄游和定居两个生态型,生殖洄游是区别两者的主要表征。为了探索嗅觉受体(OR)基因是否参与了刀鲚的生殖洄游过程,本文采用RACE技术从洄游型刀鲚中获得了mor-4k13基因,该基因全长1 098 bp,编码区长963 bp,单外显子结构,可编码320个氨基酸。预测表明,mor-4k13基因编码的蛋白质,为7个疏水性的α-螺旋跨膜结构,属G-蛋白偶联受体,有胆固醇和油酸两个配体。MOR-4K13蛋白与已报道的其他鱼类OR蛋白的同源性在40% ~ 68%之间,其中,与近缘种大西洋鲱(Clupea harengus)嗅觉受体蛋白同源性高达68%。采用Real-time PCR方法对10个组织或器官所作的荧光定量分析显示,mor-4k13基因在定居型刀鲚嗅囊和性腺中高表达,在肌肉、眼球、胃壁、肝和鳃中低表达,心肌中几乎不表达。mor-4k13基因在洄游型刀鲚嗅囊中的表达量总体高于定居型,且洄游型雄性刀鲚嗅囊中此基因的表达量约是其雌性嗅囊中的3倍。这表明mor-4k13基因不仅与嗅觉功能和性腺发育相关,也可能与生殖洄游习性相关,不同性别的个体间也存在着嗅觉能力的差异。
英文摘要:
      Japanese Grenadier Anchovy (Coilia nasus) in the Yangtze River has migratory and sedentary ecotypes, and spawning migration is a major phenotypic difference between the two groups. In order to explore whether olfactory receptor genes are involved in the process of spawning migration in C. nasus, the mor-4k13 gene encoding olfactory receptor was cloned from olfactory rosette RNA of migratory C. nasus using RACE. The mor-4k13 gene was 1 098 bp in length with an open reading frame of 963 bp which encoded 320 amino acid residues, and this gene was single exon (Fig. 2). Gene structure analysis indicated that this protein had seven hydrophobic and alpha helix transmembrane structure, belonging to the G-protein-coupled receptor superfamily (Fig. 4), and had two ligands including cholesterol and oleic acid. The homology between mor-4k13 protein of C. nasus and OR protein of other reported fishes ranged from 40% to 68%, and the homology of closely related species Atlantic Herring (Clupea harengus) was 68% (Table 2). The result based on qRT-PCR among 10 tissues and organs of the settlement population revealed that the mor-4k13 gene was expressed strongly in female olfactory rosette and ovary. Its expression in female olfactory rosette and gonad was high, but weak in muscle, eyes, stomach, liver and gills, and almost no expression was found in the heart (Fig. 5). In addition, the overall expression of the mor-4k13 gene in migratory C. nasus olfactory rosette was higher than in the sedentary group. In the migratory population, its expression in male olfactory rosette was 3 times higher than in female olfactory rosette (Fig. 6). In summary, the mor-4k13 gene not only was related to olfactory function, but also participated in gonadal development or associated with reproductive migration, and there existed gender gap in the olfactory ability.
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