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Cloning, characterization and expression analysis of a novel member of proteasomal subunits gene in turbot, Scophthalmus maximus
大菱鲆Lmp7基因克隆及表达分析
Received:August 18, 2013  Revised:April 18, 2014
DOI:
Key words:turbot ,Lmp7 ,qRT-PCR ,Tkcell
中文关键词:  大菱鲆,Lmp7 ,qRT-PCR ,Tkcell
基金项目:Finance special program of Tianjin and The transformation project of Tianjin Agricultural Achievements
Author NameAffiliationAddress
zhangbo Tianjin Bohai Sea Fisheries Research Institute 天津经济技术开发区海通街50号
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Abstract:
      The proteasome is a large, polymeric protease complex responsible for intracellular protein degradation and generation of peptides that bind to class I major histocompatibility complex (MHC) molecules. In this report we have identified a new member of proteasomal subunits from turbot (Scophthalmus maximus). The full-length cDNA sequence of turbot proteasomal subunit was obtained and sequence analysis indicated that its primary structure was highly similar to LMP7 from other vertebrates. Here we report the relationship between the turbot LMP7 expression and immune response after pathogen infection. qRT-PCR demonstrated that LMP7 was expressed differently in various tissues with higher expression in spleen, liver, muscle and skin, respectively. The LMP7 expression level was the highest at 96 h after challenge with LCDV and was the highest at 12 h after challenge with Vibrio anguillarum (V. anguillarum) in turbot liver, kidney and spleen, respectively. Furthermore, the expression of LMP7 also distinctly increased in turbot kidney cells (TK) after challenge with V. anguillarum at 24 h and LCDV at 96 h. These results indicated that the turbot LMP7 protein participates in the immune response of turbot and may play a signi?cant role in the immune process.
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。在这份报告中,我们已经确定了大菱鲆的蛋白酶体亚基的一个新成员。蛋白酶体亚基的全长cDNA序列已经获得,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。我们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在各种组织中的表达,其在脾,肝,肌肉和皮肤中,分别有不同程度的表达。 在感染牙鲆淋巴囊肿病毒96小时后,LMP7表达大菱鲆的肝脏、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。
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