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绿鳍马面鲀ERα 基因部分cDNA 序列克隆及其表达研究
赵梅琳1, 温海深1, 张冬茜1, 何 峰1, 李吉方1
中国海洋大学 水产学院
摘要:
雌二醇是与生殖相关的重要的性类固醇激素之一, 它与雌激素受体相结合作用于靶器官从而发挥生物活性。为进一步阐明绿鳍马面鲀雌激素受体α(Estrogen receptor α , ERα)的生理功能提供了科学依据, 同时也为其人工繁育提供一定的理论基础, 作者利用简并引物扩增得到绿鳍马面鲀(Navodon septentrionalis)ERα 部分cDNA 序列, 共1410bp, 编码470 个氨基酸。通过半定量RT-PCR 技术分别检测了其在雌鱼、雄鱼中各组织的表达情况, 结果表明ERα 在雌鱼体内的表达较雄鱼更广泛, 在雌鱼垂体、心脏和卵巢的表达量最高, 在雄鱼的垂体、肝脏、肾脏、精巢也有较高的表达。采用实时定量RT-PCR技术, 分析了ERα 在性腺中的周年表达情况, 卵巢中ERα mRNA 在5 月表达量最高, 7 月最低: 精巢中在5 月最低, 9 月最高。该项研究表明ERα 与绿鳍马面鲀生殖周期存在密切关联, 提示在性腺发育过程中发挥一定作用。
关键词:  绿鳍马面鲀(Navodon septentrionalis)  雌激素受体  cDNA 部分序列克隆  组织分布  mRNA 表达
DOI:10.11759/hykx20130517005
分类号:
基金项目:山东省科技攻关计划课题资助项目(2009GG10005017)
Cloning of partial cDNA of estrogen receptor α and expression analysis during the reproductive cycle in Navodon septentrionalis
Abstract:
Estrogens are critical sex steroid hormones involved in reproduction. And they need to bind to estrogen receptors (ER) in target organ for biological activity. To explore the physiological function of estrogen receptor α (ERα) in Navodon septentrionalis and provide some theoretical basis for artificial breeding, a partial cDNA sequence of N. septentrionalis ERα gene was obtained by degenerate primer PCR amplification. The length of the cDNA sequence is 1410 bp, and encodes 470 amino acids. The tissue expression analysis using semi-quantitative RT-PCR showed that it had a more restricted distribution in male fish. In female, pituitary, heart and ovary had high expression levels, and in male, pituitary, liver, kidney and testis had relatively high expression levels. In addition, we analyzed the relative expression level of ERα in reproductive cycle in ovary and testis by quantitative RT-PCR, respectively. It revealed that the highest expression level was in May, and the lowest in July in ovary, while in testis, the lowest was in May, and the highest in September. This study indicates that ERα has a strong relationship with the reproductive cycle of N. septentrionalis, suggesting ERα should play a role in gonads development.
Key words:  Navodon septentrionalis  estrogen receptor α (ERα)  partial cDNA cloning  tissue distribution  mRNA expression
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