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首页> 外文期刊>Fish Physiology and Biochemistry >Cathepsin L in the orange-spotted grouper, Epinephelus coioides: molecular cloning and gene expression after a Vibrio anguillarum challenge
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Cathepsin L in the orange-spotted grouper, Epinephelus coioides: molecular cloning and gene expression after a Vibrio anguillarum challenge

机译：橘斑石斑鱼Epinephelus coioides中的组织蛋白酶L：鳗弧菌攻击后的分子克隆和基因表达

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摘要

The orange-spotted grouper, Epinephelus coioides, is an important fish maricultured in many Asian countries. In the present study, the full-length cDNA of cathepsin L, an immunity related gene of fishes, was isolated from E. coioides using rapid amplification of cDNA ends (RACE). It is 1,443 bp in length, including an open reading frame (ORF) of 1,011 bp. The open reading frame encoded a preproprotein of 336 amino acids (aa), which consisted of a signal peptide of 16 aa, a proregion peptide of 98 aa and a mature peptide of 222 aa. The preproprotein contained an oxyanion hole (Gln), a catalytic triad formed by Cys, His and Asn, and the conserved ERWNIN, GNFD and GCNGG motifs, all characteristic of cathepsin L. Homology analysis revealed that the deduced amino acid sequence of E. coioides cathepsin L shared 80.1-94.8% identity with those of reported fishes. Tissue-dependent mRNA expression analysis showed that the cathepsin L transcript was expressed in all the examined tissues of the healthy E. coioides, being highest in the liver and moderate in the heart, gonad and intestine. After Vibrio anguillarum stimulation, the mRNA expression of cathepsin L in E. coioides was significantly increased in the skin, fin, gills, liver, blood, spleen, head kidney and intestine, with the highest observed in the spleen (10.6-fold) at 12 h post-injection and the next in blood (7.5-fold) at 8 h post-injection. These results provided initial information for further studies on the physiological and immunological roles of the cathepsin L gene in the orange-spotted grouper.Digital Object Identifier http://dx.doi.org/10.1007/s10695-012-9676-3
机译：斑纹石斑鱼石斑鱼（Epinephelus coioides）是许多亚洲国家/地区养殖的重要鱼类。在本研究中，通过快速扩增cDNA末端（RACE）从大肠杆菌中分离出组织蛋白酶L的全长cDNA，这是一种与鱼类免疫相关的基因。它的长度为1,443 bp，包括一个1,011 bp的开放阅读框（ORF）。开放阅读框编码336个氨基酸（aa）的前原蛋白，其由16个氨基酸的信号肽，98个氨基酸的前区肽和222个氨基酸的成熟肽组成。前蛋白原包含一个氧负离子孔（Gln），一个由Cys，His和Asn形成的催化三联体以及保守的ERWNIN，GNFD和GCNGG基序，都是组织蛋白酶L的特征。组织蛋白酶L与报告的鱼类具有80.1-94.8％的同一性。组织依赖性mRNA表达分析表明，组织蛋白酶L转录物在健康大肠杆菌的所有检查组织中表达，在肝脏中最高，在心脏，性腺和肠中中等。刺激鳗弧菌后，皮肤，鳍、,、肝脏，血液，脾脏，头部肾脏和肠中肠组织大肠杆菌中组织蛋白酶L的mRNA表达显着增加，在脾脏中最高（10.6倍）。注射后12小时，注射后8小时血样（7.5倍）。这些结果为进一步研究组织蛋白酶L基因在橙色斑点石斑鱼中的生理和免疫学作用提供了初步信息。数字对象标识符http://dx.doi.org/10.1007/s10695-012-9676-3

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