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《中国工程科学》 >> 2014年 第16卷 第9期

半滑舌鳎促黄体激素基因克隆和表达分析及其血清浓度测定

中国水产科学研究院黄海水产研究所,农业部海洋渔业可持续发展重点实验室,青岛市海水鱼类种子工程与生物技术重点实验室,山东青岛 266071

资助项目 :现代农业产业技术体系建设专项资金(CARS-50);国家自然科学基金项目(31201982);国家国际科技合作专项项目(2013DFA31410);山东省优秀中青年科学家科研奖励基金项目(BS2013SW042);2012 年度留学人员科技活动项目择优资助经费资助 收稿日期: 2014-06-20 发布日期: 2014-09-16 08:55:42.000

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摘要

利用RACE技术首次克隆了半滑舌鳎脑垂体中促黄体激素(LH)基因cDNA全长序列。该基因全长670 bp,开放阅读框为477 bp,编码158 个氨基酸。与其他脊椎动物的LH成熟肽氨基酸序列同源性比较表明:半滑舌鳎LH与鲽形目和鲈形目同源性高58 %~68 %。另外,半滑舌鳎LH含有12 个保守的半胱氨酸残基和1 个N-糖基化位点(19~21 NQT)。实时荧光定量组织表达分析表明,LH mRNA在脑、垂体、卵巢等组织中表达,垂体中表达量最丰富,其他组织尤以脑、性腺表达量较高,推测半滑舌鳎LH可能具有广泛的垂体外生理功能;实时荧光定量PCR方法对繁殖周期雌性半滑舌鳎LH表达水平进行测定,结果表明,LH mRNA在Ⅱ~Ⅵ各繁殖周期的脑、垂体、卵巢3 种组织都有表达,但表达水平有差异,在Ⅳ、Ⅴ期表达量最丰富,说明LH主要促进卵母细胞最终成熟及排卵。利用125I 标记的放射免疫测定(RIA)技术检测繁殖周期半滑舌鳎血清LH浓度,结果表明LH在Ⅴ期含量最丰富,也说明LH主要促进卵母细胞最终成熟及排卵。

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