| ZHANG Bo,WANG Xianli,SONG Wenping,ZHENG debin,MA Chao,XIAO Guangxia,Jia Lei.Cloning, characterization, and expression of a novel member of proteasomal subunits gene in turbot, Scophthalmus maximus[J].Acta Oceanologica Sinica,2015,34(1):110-118 |
| 大菱鲆蛋白酶体亚基LMP7基因的克隆及表达分析 |
| Cloning, characterization, and expression of a novel member of proteasomal subunits gene in turbot, Scophthalmus maximus |
| 投稿时间：2013-08-18 修订日期：2014-04-18 |
| DOI：10.1007/s13131-015-0586-y |
| 中文关键词: 大菱鲆 Lmp7 qRT-PCR Tkcell |
| 英文关键词:major histocompatibility complex (MHC) LMP7 qRT-PCR immune response turbot Vibrio anguillarum |
| 基金项目:The finance special program of Tianjin and the transformation project of Tianjin Agricultural Achievements. |
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| 中文摘要: |
| 蛋白酶体是一个大型的, 负责细胞内蛋白质降解和代谢类的家族. 它与主要组织相容性复合体(MHC)分子结合形成肽聚合物蛋白酶复杂体系从而发挥重要作用. 这份研究报告确定了大菱鲆的蛋白酶体亚基的一个新成员. 该蛋白酶体亚基的全长cdNA序列已经获得, 该cdNA含有一个117 bp的5'非编码区, 825 bp的开放阅读框(ORF)和330 bp的3'非编码区. 3'端非编码区包含在一个典型的多聚腺苷酸化终止密码信号(AATAAA), 之后是一个长24 bp的poly (A)尾巴. 通过密码子分析发现开放阅读框编码274个氨基酸. 序列分析表明, 其主要结构与其他脊椎动物的LMP7序列高度相似. 实验还同时考察了在经受不同病原体感染后, LMP7表达水平和免疫反应之间的关系. 定量RT-PCR表明LMP7在各种组织中的差异表达: 其在脾, 肝, 肌肉和皮肤中, 分别有不同程度的高表达. 在感染牙鲆淋巴囊肿病毒96小时后, LMP7表达在大菱鲆的肝脏、肾脏、脾脏达到最高. 在12 h的鳗弧菌感染过程中, 分别在大菱鲆的肝, 肾, 脾检测到LMP7的高表达. 此外, LMP7的表达在大菱鲆肾细胞(TK)感染鳗弧菌24 h和感染LCdV96h后也明显增加. 这些结果表明, 大菱鲆LMP7蛋白参与大菱鲆的免疫反应, 并可能会起到显着的免疫调节作用. |
| 英文摘要: |
| The proteasome is a large, polymeric protease complex responsible for the degradation of intracellular proteins and generation of peptides that bind to class I major histocompatibility complex (MHC) molecules. This study identified a new member of proteasomal subunits in turbots (Scophthalmus maximus). The fulllength cdNA sequence of turbot proteasomal subunit was obtained. Sequence analysis indicated that its primary structure is highly similar to that of LMP7 from other vertebrates. The relationship between the turbot LMP7 expression and immune responses to pathogen infection was reported. Quantitative reverse transcriptase polymerase chain reaction showed that LMP7 was expressed differently in various tissues, with higher expression in the spleen, liver, muscle, and skin. The LMP7 expression was the highest at 96 h after challenge with lymphocyctis disease virus (LCdV) and at 12 h after challenge with Vibrio anguillarum in the turbot liver, kidney, and spleen. Furthermore, the LMP7 expression distinctly increased in turbot kidney cells at 24 h after challenge with V.anguillarum and at 96 h after challenge with LCdV. These results indicate that the turbot LMP7 protein participates in immune responses and may play a significant role in the immune process. |
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