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

作　　者：陈学昭 张雷[1] 于珊珊[1] 毕彩红 刘春影[1] 祝茜[1] 

机构地区：[1]山东大学(威海)海洋学院,山东威海264209

出　　处：《海洋学报》2015年第10期133-140,共8页

基　　金：威海市科技攻关计划项目(0000413420608);威海市科委项目(1070432121313)

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993bp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444bp,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。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 a-mino acids.The deduced amino acid sequence had a domain called Ubcc,which was extremely conservative.The mRNA expression of TfUbcE2-D2 in healthy andVibrio 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.

关 键 词：泛素结合酶 松江鲈 基因克隆 非特异性免疫 

分 类 号：Q95[生物学—动物学]