Cloning of common carp SOCS-3 gene and its expression during embryogenesis, GH-transgene and viral infection.
Cloning of common carp SOCS-3 gene and its expression during embryogenesis, GH-transgene and viral infection.
复制标题
DOI：
10.1016/j.fsi.2009.11.012
复制
发表时间：
2010-02
影响因子：
4.7
通讯作者：
Zhang Xiao;Hong Liu;Jianjie Fu;Wei Hu;Yaping Wang;Q. Guo
中科院分区：
文献类型：
--
作者：
Zhang Xiao;Hong Liu;Jianjie Fu;Wei Hu;Yaping Wang;Q. Guo
细胞因子信号转导抑制因子3（SOCS-3）是新近发现的一个蛋白家族成员，它以负性自调节方式调节多种免疫细胞因子的应答。鲤鱼SOCS-3基因全长1603 bp，开放阅读框（ORF）为630 bp，编码209个氨基酸。鲤鱼SOCS-3分子在SRC同源序列2（SH 2）和SOCS盒中具有很高的保守性。鲤鱼SOCS-3的激酶抑制区（KIR）和ESS结构域，上游的SH 2结构域，是保守的，除了一个特定的插入（PHRYK）在KIR结构域的N-末端区域。在鲤鱼SOCS-3中还发现了三个保守的半胱氨酸（Cys-102、124和193）残基和一个额外的半胱氨酸（Cys-168）残基。鲤鱼SOCS-3基因组DNA长2015 bp，含有2个外显子和1个内含子。系统发育分析表明，鲤鱼SOCS-3序列与其它已知鱼类SOCS-3序列聚为一类，其中斑马鱼SOCS-3序列与鲤鱼SOCS-3序列最接近。RT-PCR分析表明，鲤鱼SOCS-3在受精后4 h开始表达，在胚胎发生过程中表达量逐渐增加，直至受精后4 w。RT-qPCR分析表明，鲤鱼SOCS-3基因主要在鳃、头肾、胸腺和皮肤中表达，其次是脾脏和外周血，在肾、肠、肝和肌肉中表达量较低;生长激素组SOCS-3基因在胸腺、头肾、脾脏和肠中表达量显著增加SVCV（spring viraemia of carp virus）感染后，鲤鱼SOCS-3基因在鳃、肠、胸腺、脾脏、头肾和肾脏组织中的转录水平显著上调，并呈时间依赖性。这些结果表明，硬骨鱼SOCS-3可能在调节病毒诱导的先天免疫反应和防止某些细胞因子与病毒刺激的过度作用中发挥积极作用。
As a member of a newly discovered protein family, the suppressor of cytokine signalling 3 (SOCS-3) has been shown to regulate the responses of many immune cytokines in a negative auto-regulatory manner. The full-length cDNA of common carp SOCS-3 was 1603 bp and contained a 630 bp open reading frame (ORF) coding for a protein of 209 amino acids. Carp SOCS-3 molecule was well conserved especially in the SRC homology 2 (SH2) and the SOCS box. The kinase inhibitory region (KIR) and ESS domains, upstream of the SH2 domain, were conserved in carp SOCS-3, except for a specific insertion (PHRYK) in the KIR domain at the N-terminal region. Three conserved cysteine (Cys-102, 124 and 193) residues, and one additional cysteine (Cys-168) residue, were also found in carp SOCS-3. The 2015 bp genomic DNA of carp SOCS-3 contained two exons and one intron. Phylogenetic analysis showed that carp SOCS-3 sequence grouped with other known fish SOCS-3 sequences with zebrafish SOCS-3 as the closest neighbour. RT-PCR analysis showed that carp SOCS-3 was initially expressed at 4 h pf (post-fertilization) and gradually increased up to 4 w pf during embryogenesis. By RT-qPCR analysis, carp SOCS-3 gene was predominantly detected in gill, head kidney, thymus and skin, followed by spleen and peripheral blood, lower expression level was detected in kidney, intestine, liver and muscle; the SOCS-3 transcript was significantly increased in thymus, head kidney, spleen and intestine of GH (growth hormone)-transgenic carp; after SVCV (spring viraemia of carp virus) infection, the carp SOCS-3 transcript was significantly up-regulated in gill, intestine, thymus, spleen, head kidney and kidney tissues in a time-dependent manner. These results suggest that teleost SOCS-3 may play an active role in the modulation of viral-induced innate immune response and in preventing the overaction of some cytokines with viral stimulation.