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光倒刺鲃mc5r基因的克隆及对饥饿的响应

周惠强 陈凯棱 舒琥 杨扬 钟东明 张明清

引用本文:
Citation:

光倒刺鲃mc5r基因的克隆及对饥饿的响应

    作者简介: 周惠强(1993—),男,硕士研究生,从事鱼类生理与分子生物学研究。E-mail: 852167088@qq.com;
    通讯作者: 舒琥, shuhu001@126.com
  • 中图分类号: S 917.4

Cloning and starvation response of mc5r gene in Spinibarbus hollandi

    Corresponding author: Hu SHU, shuhu001@126.com ;
  • CLC number: S 917.4

  • 摘要: 通过同源克隆和染色体步移对光倒刺鲃 (Spinibarbus hollandi)黑素皮质素受体-5 (melanocortin receptor-5,mc5r)基因进行了克隆,应用生物信息学方法对mc5r基因序列和推测的氨基酸序列进行分析,采用荧光定量PCR技术研究其mRNA的组织分布、日周期变化以及饥饿再投喂的表达情况。结果表明mc5r基因长度为1 947 bp,含有1个987 bp的外显子,编码329个氨基酸。通过序列同源比对及系统发育进化分析,光倒刺鲃mc5r的氨基酸序列与鲤 (Cyprinus carpio)、犀角金线鲅 (Sinocyclocheilus rhinocerous)的mc5r同源性达到98%。光倒刺鲃mc5r在大脑中显著表达,在间脑、心脏的表达量次之,在中脑、小脑表达量较少,在肾脏、胃、肌肉、脾、肠、性腺、肝脏、延脑、眼中有微量表达。光倒刺鲃mc5r mRNA的日周期表达量随昼夜节律而变化,饥饿再投喂实验发现饥饿再投喂组鱼其脑中表达量会显著升高,随后会有下降趋势,表明mc5r可能与光倒刺鲃的摄食调控有关。
  • 图 1  光倒刺鲃mc5r基因组扩增序列结果

    Figure 1.  Sequence results of S. hollandi mc5r genome DNA

    图 2  光倒刺鲃mc5r核苷酸序列以及推测氨基酸序列

    Figure 2.  mc5r nucleotide sequence and deduced amino acid sequence of S. hollandi

    图 3  光倒刺鲃与其他物种mc5r氨基酸序列构建的系统发育树

    Figure 3.  Phylogenetic tree constructed from mc5r amino acid sequence of S. hollandi and other species

    图 4  光倒刺鲃mc5r在不同组织中的表达

    Figure 4.  Expression of S. hollandi mc5r in different tissues

    图 5  光倒刺鲃大脑mc5r的日周期表达

    Figure 5.  Daily periodicitye expression of mc5r in brain of S. hollandi

    图 6  光倒刺鲃mc5r mRNA饥饿再投喂的表达情况

    Figure 6.  Expression profiles of mc5r at starvation and refeeding condition in S. hollandi

    表 1  光倒刺鲃mc5r基因克隆和表达分析的引物

    Table 1.  Primers for cloning and expression analysis of mc5r gene in S. hollandi

    引物
    primer
    引物序列 (5'−3')
    primer sequence
    用途
    application
    产物大小/bp
    product size
    F1AAGATGCCCTGTCTACCAACCC克隆850
    R1GGTCCAAATCCCTCCAATGATG克隆
    F2GGAGGGAAAACAACAGCACAAGC克隆750
    R2GGTCCAAATCCCTCCAATGATG克隆800
    F3GGTTAAAGATGCCCAACCCACC克隆800
    R3GGATGAGGTGGAGAAAGAACGG克隆800
    F4CTACTTACTCACCAACCGCCAG克隆850
    R4CTTTGATCGACAGCGTGTCCC克隆900
    FAACACTTCAGAGGCCACCCTGqRT-PCR150
    RTGAGCTGCTCGCATGCTTTGGqRT-PCR
    18sFTGATCGTCTTCGAGCCTCTGAC内参150
    18sRAATTCTTGGACCGGCGCAAGAC内参
    下载: 导出CSV
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光倒刺鲃mc5r基因的克隆及对饥饿的响应

    作者简介:周惠强(1993—),男,硕士研究生,从事鱼类生理与分子生物学研究。E-mail: 852167088@qq.com
    通讯作者: 舒琥, shuhu001@126.com
  • 广州大学生命科学学院,广东广州 510006

摘要: 通过同源克隆和染色体步移对光倒刺鲃 (Spinibarbus hollandi)黑素皮质素受体-5 (melanocortin receptor-5,mc5r)基因进行了克隆,应用生物信息学方法对mc5r基因序列和推测的氨基酸序列进行分析,采用荧光定量PCR技术研究其mRNA的组织分布、日周期变化以及饥饿再投喂的表达情况。结果表明mc5r基因长度为1 947 bp,含有1个987 bp的外显子,编码329个氨基酸。通过序列同源比对及系统发育进化分析,光倒刺鲃mc5r的氨基酸序列与鲤 (Cyprinus carpio)、犀角金线鲅 (Sinocyclocheilus rhinocerous)的mc5r同源性达到98%。光倒刺鲃mc5r在大脑中显著表达,在间脑、心脏的表达量次之,在中脑、小脑表达量较少,在肾脏、胃、肌肉、脾、肠、性腺、肝脏、延脑、眼中有微量表达。光倒刺鲃mc5r mRNA的日周期表达量随昼夜节律而变化,饥饿再投喂实验发现饥饿再投喂组鱼其脑中表达量会显著升高,随后会有下降趋势,表明mc5r可能与光倒刺鲃的摄食调控有关。

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