刀鲚MOR-2AK2的克隆、序列分析及组织表达
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上海海洋大学鱼类研究室,上海海洋大学鱼类研究室,上海海洋大学鱼类研究室,上海海洋大学鱼类研究室

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公益性行业(农业)科研专项(201203065);国家自然科学基金(31172407,31472280);高等学校博士学科点科研专项(20123104110006);上海高校水产学高峰学科


Cloning, sequence analysis and tissue expression of Coilia nasus olfactory receptor gene MOR-2AK2
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Laboratory of Ichthyology,Shanghai Ocean University,Laboratory of Ichthyology,Shanghai Ocean University,Laboratory of Ichthyology,Shanghai Ocean University,Laboratory of Ichthyology,Shanghai Ocean University

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    摘要:

    嗅觉是鱼类感知周围环境的重要工具,可能参与生殖洄游的过程。嗅觉由嗅觉受体(olfactory receptor)基因所编码的受体蛋白识别气味分子所引发,主嗅觉受体(MOR)基因是数量最大的一类嗅觉基因,可识别水溶性气味分子。为了弄清刀鲚定居型与洄游型种群的主嗅觉基因差异,本研究通过RACE技术获得了洄游型刀鲚MOR-2AK2基因,其开放阅读框长度972 bp,为单外显子结构,可编码323个氨基酸残基。预测表明,MOR-2AK2基因所编码的蛋白为7个疏水性的α-螺旋跨膜结构,属于G-蛋白偶联受体。对10种组织所作的定量分析表明,MOR-2AK2基因在嗅囊和性腺中的表达量远远高于其他组织器官,并且嗅囊中的表达量还高于性腺中的7~25倍。MOR-2AK2基因的表达量也存在性别差异,其中雌性嗅囊中的表达量约为雄性中的2倍,但精巢中的表达量却约是卵巢中的2倍。序列分析显示,MOR-2AK2基因的5'-UTR区域存在着一段微卫星序列(GT)5,其中定居型多出洄游型14个碱基(GTGTGTGTGTGTTT),这导致了二者所编码的氨基酸序列的相似度仅为84%。这些结果表明,MOR-2AK2基因不但与嗅觉功能有关,也可能参与了刀鲚的性腺发育或生殖洄游过程,同时也可能与定居型种群的形成相关。

    Abstract:

    Olfaction is the important tool of sensing their surroundings to fish and may take part in spawning migration. Fish perform olfactory function by the means of the olfactory receptor proteins that olfactory receptor genes encode. The main olfactory receptor gene is the largest number of olfactory receptor families, which can identify water-soluble odor molecules. To find out the main olfactory receptor genes' differences of C. nasus between settlement population and migratory population, the MOR-2AK2 cDNA sequence of migratory C. nasus using rapid amplification of cDNA ends (RACE) was cloned. The sequencing results showed that the open reading frame was 972 bp in length, encoding 323 amino acid residues. The MOR-2AK2 protein had seven hydrophobic and alpha helix transmembrane structures, belonging to the G-protein-coupled receptor superfamily. Based on qRT-PCR among 10 tissues in migratory C. nasus, the result revealed that MOR-2AK2 was expressed in the olfactory rosettes and sex glands more highly than in others. Further analysis showed, the expression level in olfactory rosettes was 5 to 27 times higher than that in sex glands. In addition, there were differences in tissue expression quantity between male and female. The female olfactory rosette's expression level was 2 times that in male olfactory rosette. But the testis' expression level was 2 times that in ovary. Sequence analysis displayed that the 5 untranslated regions of MOR-2AK2 gene had microsatellite sequence, (GT)5. The settlement population had a 14 bp more sequence (GTGTGTGTGTGTTT) than the migratory population. And the amino acid sequence similarity was just 84% between them. From the above, MOR-2AK2 gene not only took part in olfactory function, but also participated in gonad development or breeding migratory, and may be involved in the formation of settlement population.

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王聪,朱国利,唐文乔,刘东,杨金权.刀鲚MOR-2AK2的克隆、序列分析及组织表达[J].水产学报,2016,40(1):11~23

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  • 收稿日期:2015-01-29
  • 最后修改日期:2015-11-03
  • 录用日期:2016-01-27
  • 在线发布日期: 2016-01-27
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