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从大西洋鳕鱼(Gadus morhua)中分离和鉴定两个编码转谷氨酰胺酶的 cDNA。

Isolation and characterisation of two cDNAs encoding transglutaminase from Atlantic cod (Gadus morhua).

机构信息

Centre for Organelle Research, Faculty of Science and Technology, University of Stavanger, 4021 Stavanger, Norway.

Nofima, Box 6122, N-9291 Tromsø, Norway.

出版信息

Fish Shellfish Immunol. 2014 Jan;36(1):276-83. doi: 10.1016/j.fsi.2013.11.014. Epub 2013 Dec 3.

Abstract

Two cDNAs encoding transglutaminase (TG) were identified in a subtractive cDNA library prepared from the head kidney of poly I:C stimulated Atlantic cod (Gadus morhua). Full-length TG-1 and TG-2 cDNA were cloned from the head kidney by a reverse-transcription polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE). The deduced amino acid (aa) sequence for TG-1 was 695 aa with an estimated molecular mass of 78.3 kDa, while TG-2 was a 698 aa protein with an estimated molecular mass of 78.8 kDa. The two proteins were named TG-1 and TG-2 and both possess transglutaminase/protease-like homologous domains (TGc) and full conservation of amino acids cysteine, histidine, and aspartate residues that form the catalytic triad. Sequence analysis showed high similarity (93.1%) with Alaska pollock TG, and the TGs were grouped together with TGs from chum salmon, Japanese flounder, Nile tilapia, and red sea bream in addition to Alaska pollock in phylogenetic analysis. Interestingly, they showed different tissue distribution with highest constitutive expression in reproductive and immunological organs, indicating important roles in these organs. Furthermore, the up-regulation of TG-1 and TG-2 in head kidney after stimulating Atlantic cod with poly I:C suggested a role of TGs in immune response in Atlantic cod.

摘要

从聚肌胞刺激的大西洋鳕鱼头肾制备的消减 cDNA 文库中鉴定出两种编码转谷氨酰胺酶 (TG) 的 cDNA。通过逆转录聚合酶链反应 (RT-PCR) 和 cDNA 末端快速扩增 (RACE) 从头肾中克隆出全长 TG-1 和 TG-2 cDNA。TG-1 的推导氨基酸 (aa) 序列为 695 aa,估计分子量为 78.3 kDa,而 TG-2 是一种 698 aa 蛋白质,估计分子量为 78.8 kDa。这两种蛋白质分别命名为 TG-1 和 TG-2,均具有转谷氨酰胺酶/蛋白酶样同源结构域 (TGc),并且半胱氨酸、组氨酸和天冬氨酸残基完全保守,这些残基构成催化三联体。序列分析显示与阿拉斯加狭鳕 TG 具有高度相似性 (93.1%),并且在系统发育分析中,TGs 与大麻哈鱼、日本牙鲆、尼罗罗非鱼和真鲷的 TG 一起与阿拉斯加狭鳕分组。有趣的是,它们表现出不同的组织分布,在生殖和免疫器官中具有最高的组成型表达,表明在这些器官中具有重要作用。此外,用聚肌胞刺激大西洋鳕鱼后头肾中 TG-1 和 TG-2 的上调表明 TG 在大西洋鳕鱼的免疫反应中起作用。

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