半滑舌鳎孕酮受体膜组分1基因的克隆及组织和时空表达规律
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1. 大连海洋大学, 水产与生命学院, 辽宁 大连 116023;
2. 农业部海洋渔业可持续发展重点实验室, 中国水产科学研究院 黄海水产研究所, 山东 青岛 266071;
3. 青岛海洋科学与技术国家实验室, 海洋渔业科学与食物产出过程功能实验室, 山东 青岛 266237

作者简介:

张金勇(1987-),男,硕士,研究方向为鱼类繁殖内分泌调控机制研究.E-mail:jinyongzhang2013@126.com

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S917

基金项目:

国家自然科学基金项目(31201982);山东省优秀中青年科学家科研奖励基金项目(BS2013SW042);国家鲆鲽类产业技术体系项目(CARS-50).


Molecular cloning of PGRMC1 from half-smooth tongue sole (Cyno-glossus semilaevis) and its tissue and spatio-temporal expression patterns
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Affiliation:

1. College of Fisheries and Life Science, Dalian Ocean University, Dalian 116023, China;
2. Key Laboratory of Sustainable Development of Marine Fisheries, Ministry of Agriculture;Yellow Sea Fisheries Re-search Institute, Chinese Academy of Fishery Scie

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    摘要:

    运用同源克隆和RACE方法获得了半滑舌鳎( cDNA序列全长1335 bp,开放阅读框长546 bp;其编码的蛋白是单次跨膜蛋白,在N端13~35位氨基酸残基处有一个跨膜区域。半滑舌鳎PGRMC1氨基酸序列与青鳉()相似度为81.2%。系统进化分析表明,半滑舌鳎PGRMC1与鳉形目和鲀形目鱼类PGRMC1聚为一个分支。采用实时荧光定量RT-PCR技术研究发现,性成熟雌性半滑舌鳎PGRMC1 mRNA在脑、垂体和卵巢中的周期表达变化特征显示:脑中 mRNA表达量在卵巢发育V期,达峰值(<0.05)。综上,半滑舌鳎基因主要参与卵巢的发育、和成熟过程,并在繁殖期介导孕酮调控卵母细胞的最终成熟,研究结果为半滑舌鳎繁殖内分泌调控研究提供了基础资料。

    Abstract:

    To investigate the role of progesterone receptor membrane component 1 (PGRMC1) in oocyte maturation in the half-smooth tongue sole (PGRMC1 from the ovary using homology cloning and rapid amplification of cDNA ends methods. The full-length cDNA of was 1335 bp long with an open reading frame of 546 bp encoding a 181 amino acid preprohormone with a deduced molecular mass of 20.64 kDa and a theoretical isoelectric point of 4.67. The precursor was a single transmembrane protein with an N-terminal transmembrane domain. The predicted transmembrane domain was located at positions 13-35 of the deduced PGRMC1 protein. Sequence alignment of the PGRMC1 precursor protein with corresponding sequences from other species revealed that the highest identity (82.87%) was with the PGRMC1 from (81.22%). In a phylogenetic analysis, the PGRMC1 clustered with its counterparts in the Cyprinodontiformes and Tetraodontiformes. The transcript levels of were high in the ovary, moderate in the liver and brain, and low in other tissues. We also monitored changes in transcript levels in the pituitary, brain, and ovary at different ovarian developmental stages. The mRNA levels increased sharply in the brain at stage Ⅲ, remained at high levels until stage V, and then decreased at stage VI. In the pituitary, the transcript levels of increased to peak at stage V and decreased significantly after ovulation (stage VI). Similarly, in the ovary, the levels increased gradually to peak at stage V and then decreased markedly after spawning (stage VI). Taken together, our results shed light on the role of PGRMC1 in oocyte maturation and in regulating reproductive endocrine function in .

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张金勇,柳学周,史宝,徐永江,李晓妮,常亚青,王滨.半滑舌鳎孕酮受体膜组分1基因的克隆及组织和时空表达规律[J].中国水产科学,2016,23(5):1080-1090
ZHANG Jinyong, LIU Xuezhou, SHI Bao, XU Yongjiang, LI Xiaoni, CHANG Yaqing, WANG Bin. Molecular cloning of PGRMC1 from half-smooth tongue sole (Cyno-glossus semilaevis) and its tissue and spatio-temporal expression patterns[J]. Journal of Fishery Sciences of China,2016,23(5):1080-1090

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  • 在线发布日期: 2016-09-05
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