| 曼氏无针乌贼Phb2基因的克隆和其在不同生长阶段的表达分析 |
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| 引用本文： | 毋玉婷,郭宝英,祁鹏志,吕振明,吴常文,史会来,平洪领.曼氏无针乌贼Phb2基因的克隆和其在不同生长阶段的表达分析[J].中国水产科学,2018,25(1):9-17. |
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| 作者姓名： | 毋玉婷 郭宝英 祁鹏志 吕振明 吴常文 史会来 平洪领 |
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| 作者单位： | 浙江海洋大学国家海洋设施养殖工程技术研究中心;浙江省海水增养殖重点实验室浙江省海洋水产研究所; |
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| 基金项目： | 科技部国际科技合作港澳台项目（2014DFT30120）;农业部渔业渔政管理局物种资源保护项目（17162130135252089）;舟山市科技局项目（2013C41001）;浙江海洋大学"海洋科学"省重中之重学科开放课题（20160109）;浙江省科技计划项目（2015F50055） |
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| 摘 要： | Phb2是抗增殖蛋白(Prohibitin)的一个亚型。本研究利用RACE技术首次获得了曼氏无针乌贼(Sepiella japonica)Phb2基因的全长序列,该序列全长为1283 bp,其中5′非编码区(5′-UTR)150 bp,3′非编码区(3′-UTR)236 bp,开放阅读框(ORF)897 bp,预测编码298个氨基酸。序列分析表明,曼氏无针乌贼Phb2基因编码的蛋白无信号肽,第20~42位氨基酸为跨膜结构,包含1个PHB家族的结构域。采用荧光定量PCR技术检测其在曼氏无针乌贼不同生长时期(幼体、产卵前、产卵中、产卵后濒死前)各组织中的表达情况,结果显示,Phb2在4个时期的各组织中均有表达,其中,幼体和濒死前表达量较低,产卵前和产卵中的表达量则相对较高,在各时期中脑、肝表达较多。视腺作为与生殖调控有关的内分泌器官,其Phb2基因表达具有时期差异性,在产卵后的表达量由产卵时期的高表达大幅下降,说明Phb2具有周期调节、控制细胞衰老和凋亡的多效性功能。本研究结果可为进一步了解抗增殖蛋白对曼氏无针乌贼的抗衰老作用提供参考。
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| 关 键 词： | 抗增殖蛋白Phb2 表达分析 曼氏无针乌贼 抗衰老 |
| 修稿时间： | 2018/1/19 0:00:00 |
Cloning of the Phb2 gene and analysis of its developmental stage distribution in Sepiella japonica |
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| WU Yuting,GUO Baoying,QI Pengzhi,LYU Zhenming,WU Changwen,SHI Huilai,PING Hongling.Cloning of the Phb2 gene and analysis of its developmental stage distribution in Sepiella japonica[J].Journal of Fishery Sciences of China,2018,25(1):9-17. |
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| Authors: | WU Yuting GUO Baoying QI Pengzhi LYU Zhenming WU Changwen SHI Huilai PING Hongling |
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| Institution: | 1. National Engineering Research Center for Marine Aquaculture, Zhejiang Ocean University, Zhoushan 316022, China;2. Zhejiang Province Key Lab of Mariculture and Enhancemenl, Marine Fisheries Research Institute of Zhejiang, ZhouShan 316021, China |
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| Abstract: | Aging and death are very complex biological phenomena. In recent years, aging has become an important research topic. Researchers have explored the mechanisms of aging at the cellular and molecular levels. Sepiella japonica is widely distributed in China''s coastal areas, with high food, medicinal, and economic value; it is an important cephalopod of economic value in China. It usually dies after spawning, which is a common characteristic of cephalopods. However, the lifecycle and aging-related mechanisms in this squid are still unknown. In this study, we used S. japonica to examine prohibitin, which is a highly conserved protein that is widely present in eukaryotes from yeasts to mammals and plays an important role in cell cycle management, differentiation, aging, and anti-proliferative activity. Phb2 is a subtype of Prohibitin. We first acquired the Phb2 gene using rapid amplification of the cDNA end (RACE) technique. The full length of the sequence was 1283 bp, containing a 5''-untranslated region (5''-UTR) of 150 bp, a 3''-untranslated region (3''-UTR) of 236 bp, and an open reading frame (ORF) of 897 bp that was predicted to encode 298 amino acids. Sequence analysis showed that the protein encoded by the Phb2 gene did not contain a signal peptide, but contained a domain of the PHB family, and the fragment including 20aa to 42aa was a transmembrane structure. The sequence was 87% similar to Octopus bimaculoides. Phylogenetic analysis showed that S. japonica is clustered with mollusks and arthropods, and the chordate is another, which is in accordance with traditional taxonomy. The expression of Phb2 was detected using quantitative real-time PCR on tissues of S. japonica at different growth stages (larvae, spawning, spawning-and-postpartum). The results showed that the Phb2 gene was expressed in all four developmental stages. The expressions were lower in the larvae and spawning-and-postpartum stages and relatively high during spawning and spawning. Expressions in brain and liver tissues were high at all stages. The optic gland, an endocrine organ, was associated with reproductive regulation, and its Phb2 gene expression showed variation over time. Expression of the Phb2 gene was significantly decreased after spawning, which indicated that Phb2 had pleiotropic functions of cycle regulation, controlling cell senescence, and apoptosis. The results of this study provide a basis for further study on the anti-aging effects of Prohibitin2 in S. japonica and its expression in cephalopods, and also provides some theoretical basis and information for the causes of senility and senescence mechanisms in marine cephalopods. |
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| Keywords: | anti-proliferation protein of Phb2 expression analysis Sepiella japonica anti-aging function |
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