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陈学昭,张雷,于珊珊,毕彩红,刘春影,祝茜.松江鲈(Trachidermus fasciatus)泛素结合酶E2-D2基因的分子克隆及组织表达分析[J].海洋学报,2015,37(10):133-140
松江鲈(Trachidermus fasciatus)泛素结合酶E2-D2基因的分子克隆及组织表达分析
Molecular cloning and expression analysis of ubiquitin-conjugating enzyme E2-D2 in roughskin sculpin,Trachidermus Fasciatus
投稿时间:2015-03-25  
DOI:10.3969/j.issn.0253-4193.2015.10.013
中文关键词:  泛素结合酶  松江鲈  基因克隆  非特异性免疫
英文关键词:ubiquitin-conjugating enzyme  Trachidermus fasciatus  gene clone  innate immunity
基金项目:威海市科技攻关计划项目(0000413420608); 威海市科委项目(1070432121313)。
作者单位E-mail
陈学昭 山东大学(威海) 海洋学院, 山东 威海 264209  
张雷 山东大学(威海) 海洋学院, 山东 威海 264209  
于珊珊 山东大学(威海) 海洋学院, 山东 威海 264209  
毕彩红 山东大学(威海) 海洋学院, 山东 威海 264209  
刘春影 山东大学(威海) 海洋学院, 山东 威海 264209  
祝茜 山东大学(威海) 海洋学院, 山东 威海 264209 qianzhu@sdu.edu.cn 
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中文摘要:
      泛素结合酶(UbcE2)是蛋白泛素化过程中所必需的酶,在泛素转移和底物的特异性识别方面发挥着重要的作用。本实验利用RACE技术克隆获得了全长为993 bp的松江鲈泛素结合酶E2-D2基因cDNA序列(命名为TfUbcE2-D2),其开放阅读框为444 bp,编码147个氨基酸。通过SMART预测得知,TfUbcE2-D2含有一个UBCc结构域。同源比对结果表明该基因与其他物种的同源性为92.31%。实时荧光定量PCR显示,TfUbcE2-D2广泛表达于松江鲈各组织,在肾脏中的相对表达量最高,其次为鳃。鳗弧菌(Vibrio anguillarum)刺激后,TfUbcE2-D2在血液、脾脏、肝脏和鳃中表达均上调,其中,脾脏和鳃中的表达量上调约40倍。由以上实验结果推测,TfUbcE2-D2可能参与松江鲈的先天免疫防御。
英文摘要:
      Ubiquitin-conjugating enzyme is the second enzyme in the process of protein ubiquitination and plays an important role in ubiquitin transferring and substrate specific recognition. In this study,we cloned the full-length cDNA from the Roughskin sculpin,Trachidermus fasciatus by RACE,and named as TfUbcE2-D2. The cDNA of TfUbcE2-D2 is 993 bp,which contained an open reading frame (ORF) of 444 bp encoding a polypeptide of 147 amino acids. The deduced amino acid sequence had a domain called Ubcc,which was extremely conservative. The mRNA expression of TfUbcE2-D2 in healthy and Vibrio anguillarum challenge was detected by quantitative real-time PCR,results showed that TfUbcE2-D2 was expressed in all the detected tissues,mainly expressed in kidney and gill. The up-regulated of TfUbcE2-D2 post challenge with Vibrio anguillarum in blood,spleen,liver and gill indicated that it might be involved in the innate response of Roughskin sculpin.
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