Key Laboratory of Marine Bio-resources Sustainable Utilization, South China Sea Institute of Oceanology, Chinese Academy of Sciences, 164 West Xingang Road, Guangzhou 510301, PR China.
Fish Shellfish Immunol. 2011 Oct;31(4):600-5. doi: 10.1016/j.fsi.2011.07.002. Epub 2011 Jul 13.
Leukocyte cell-derived chemotaxin-2 (LECT2) is a multifunctional protein involved in cell growth, differentiation and autoimmunity. In this study, a new leukocyte cell-derived chemotaxin-2 (EcLECT2) gene was cloned from grouper, Epinephelus coioides, by rapid amplification of cDNA ends (RACE) PCR. The full-length cDNA sequence of EcLECT2 was 595 bp in size, containing a 5'-untranslated region (UTR) of 44 bp and a 3'-UTR of 83 bp. The deduced protein sequence of the open reading frame (465 bp) showed highest similarity (81%) to the LECT2 of the fresh-water fish Larimichthys crocea. An abundant transcription of the determined EcLECT2 mRNA has been detected in liver and skin of grouper, E. coioides. Furthermore, the expression of EcLECT2 was differentially up-regulated in liver after infection with Staphyloccocus aureus, Vibrio vulnificus, Vibrio parahaemolyticus, Saccharomyces cerevisiae and Singapore grouper iridovirus (SGIV), while the expression was down-regulated after stimulation with Concanavalin A (Con A). Recombinant mature EcLECT2 (rEcLECT2) was successfully expressed in Escherichia coli BL21 (DE3), and the antiserum against EcLECT2 was obtained for further investigations. EcLECT2 may be an important molecule in the innate immunity of grouper.
白细胞细胞衍生趋化因子-2（LECT2）是一种多功能蛋白，参与细胞生长、分化和自身免疫。在这项研究中，我们通过快速扩增 cDNA 末端（RACE）PCR 从石斑鱼（Epinephelus coioides）中克隆出一种新的白细胞细胞衍生趋化因子-2（EcLECT2）基因。EcLECT2 的全长 cDNA 序列大小为 595bp，包含 44bp 的 5'-非翻译区（UTR）和 83bp 的 3'-UTR。开放阅读框（465bp）的推导蛋白序列与淡水鱼Larimichthys crocea 的 LECT2 具有最高的相似性（81%）。在石斑鱼肝脏和皮肤中检测到 EcLECT2 mRNA 的大量转录。此外，EcLECT2 在感染金黄色葡萄球菌、创伤弧菌、副溶血弧菌、酿酒酵母和新加坡石斑鱼虹彩病毒（SGIV）后在肝脏中的表达水平差异上调，而在 Concanavalin A（Con A）刺激后表达水平下调。重组成熟 EcLECT2（rEcLECT2）在大肠杆菌 BL21（DE3）中成功表达，并获得了针对 EcLECT2 的抗血清，以进行进一步研究。EcLECT2 可能是石斑鱼先天免疫中的一个重要分子。
