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脊尾白虾(Exopalaemon carinicauda)Na⁺/K⁺-ATP酶α亚基基因在应对盐度胁迫中的作用

The roles of Na⁺/K⁺-ATPase α-subunit gene from the ridgetail white prawn Exopalaemon carinicauda in response to salinity stresses.

作者信息

Li Jitao, Ma Peng, Liu Ping, Chen Ping, Li Jian

机构信息

Key Laboratory for Sustainable Utilization of Marine Fisheries Resources, Ministry of Agriculture, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, 106 Nanjing Road, Qingdao, 266071 PR China.

Key Laboratory for Sustainable Utilization of Marine Fisheries Resources, Ministry of Agriculture, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, 106 Nanjing Road, Qingdao, 266071 PR China.

出版信息

Fish Shellfish Immunol. 2015 Feb;42(2):264-71. doi: 10.1016/j.fsi.2014.10.043. Epub 2014 Nov 8.

Abstract

Na(+)/K(+)-ATPase (NAK) is one important transporter protein and plays a key role in maintaining osmotic homeostasis in low and high salinity acclimation in variety of crustacean species. The ridgetail white prawn Exopalaemon carinicauda is an euryhaline and economic shrimp species in China, but it remains unclear about its mechanism of salinity adaption. In this study, a full-length of Na(+)/K(+)-ATPase α-subunit (α-NAK) cDNA was cloned from E. carinicauda by using rapid amplification of cDNA ends (RACE) approaches. The full-length cDNA of α-NAK was of 3680 bp, containing an open reading frame (ORF) of 3030 bp encoding a polypeptide of 1009 amino acids with the predicted molecular weight of 112.27 kDa. Eight transmembrane domains and two sites of phosphorylation and ATP binding were identified in E. carinicauda α-NAK. BLAST analysis revealed that the sequence of α-NAK amino acids of E. carinicauda shared more than 75% homologies with those of other crustacean. Real time quantitative RT-PCR analysis indicated that E. carinicauda α-NAK gene could be detected in all the tested tissues with highest expression level in gill. The expression profiles of E. carinicauda α-NAK transcripts were analyzed in gill and hepatopancreas tissues after salinity stresses. The results showed that the expression level of E. carinicauda α-NAK gene in both gill and hepatopancreas reached peak at different time after low and high salinity stresses, and showed different expression profiles. The expression profiles of proPO transcripts in gills after salinity stresses also indicated α-NAK and proPO played synergistic actions for salinity responses in E. carinicauda. These results indicated that E. carinicauda α-NAK involved in stress responses against salinity.

摘要

钠钾ATP酶(NAK)是一种重要的转运蛋白,在多种甲壳类动物适应低盐和高盐环境以维持渗透平衡的过程中发挥关键作用。脊尾白虾是中国一种广盐性的经济虾类,但其盐度适应机制尚不清楚。在本研究中,采用cDNA末端快速扩增(RACE)技术从脊尾白虾中克隆了钠钾ATP酶α亚基(α-NAK)的全长cDNA。α-NAK的全长cDNA为3680 bp,包含一个3030 bp的开放阅读框(ORF),编码一个由1009个氨基酸组成的多肽,预测分子量为112.27 kDa。在脊尾白虾α-NAK中鉴定出8个跨膜结构域以及两个磷酸化和ATP结合位点。BLAST分析表明,脊尾白虾α-NAK氨基酸序列与其他甲壳类动物的同源性超过75%。实时定量RT-PCR分析表明,在所有检测组织中均能检测到脊尾白虾α-NAK基因,其中鳃中的表达水平最高。分析了盐度胁迫后脊尾白虾鳃和肝胰腺组织中α-NAK转录本的表达谱。结果表明,在低盐和高盐胁迫后的不同时间,脊尾白虾鳃和肝胰腺中α-NAK基因的表达水平均达到峰值,且呈现出不同的表达谱。盐度胁迫后鳃中前酚氧化酶(proPO)转录本的表达谱也表明,α-NAK和proPO在脊尾白虾对盐度的响应中发挥协同作用。这些结果表明,脊尾白虾α-NAK参与了对盐度胁迫的应激反应。

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