中国水产科学2018，Vol.25Issue(1)：9-17,9.DOI:10.3724/SP.J.1118.2018.17101
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
Cloning of the Phb2 gene and analysis of its developmental stage dis-tribution in Sepiella japonica
摘要
Abstract
Aging and death are very complex biological phenomena. In recent years, aging has become an im-portant 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 char-acteristic 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 ampli-fication 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 an-other, 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 low-er in the larvae and spawning-and-postpartum stages and relatively high during spawning and spawning. Expres-sions 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 regula-tion, 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 theoret-ical basis and information for the causes of senility and senescence mechanisms in marine cephalopods.关键词
抗增殖蛋白Phb2/表达分析/曼氏无针乌贼/抗衰老Key words
anti-proliferation protein of Phb2/expression analysis/Sepiella japonica/anti-aging function分类
农业科技引用本文复制引用
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):9-17,9.基金项目
科技部国际科技合作港澳台项目(2014DFT30120) （2014DFT30120）
农业部渔业渔政管理局物种资源保护项目(17162130135252089) （17162130135252089）
舟山市科技局项目(2013C41001) （2013C41001）
浙江海洋大学"海洋科学"省重中之重学科开放课题(20160109) （20160109）
浙江省科技计划项目(2015F50055) （2015F50055）
