猪SOCS3基因的克隆、生物信息分析及热应激条件下的表达

Cloning and Bioinformatics Analysis of Porcine SOCS3 cDNA and Its Expression Pattern during Heat Stress

鉴于细胞因子信号传导抑制因子3(SOCS3)与多种疫病的发生密切相关,推测其可能在热应激致病中发挥重要作用。本研究克隆了猪SOCS3基因cDNA序列,发现SOCS3基因开放阅读框(ORF)长690bp,编码229个氨基酸的前体蛋白。其与八眉猪、马、牛、人和小鼠的氨基酸序列一致性分别为98.2%、97.4%、96.5%、97.8%和96.4%。该蛋白分子量25.1ku,等电点(PI)8.84,具有SOCS家族特异的中央SH2区、C末端40个氨基酸的SOCS box区及12个氨基酸的KIR结构区。荧光定量PCR结果显示,肠系膜淋巴结中SOCS3mRNA在热应激的第3、6和9天显著下调(P≤0.05)。但在胸腺和小肠黏膜中,尽管在第3天呈下降趋势,但在第6和第9天显著上调(P≤0.05)。本研究成功克隆了猪SOCS3基因,发现其在热应激条件下表达量发生明显改变,并呈组织和时间依赖性。

Suppressor of cytokine signaling 3(SOCS3)associated with a lot of auto-immune diseases,so it was speculated that it might play an important role in the pathological process of heat stress in pigs.The porcine SOCS3 gene was cloned in this study,it consisted of 690 bp of ORF encoding aprotein precursor with 229 amino acids,which shared high amino acid sequence identity with that of Bamei pig(98.2%),Equus caballus(97.4%),Bos Taurus(96.5%),Homo sapiens(97.8%)and Mus musculus(96.4%).The molecular weight of SOCS3 protein was 25.1ku,and PI was 8.84.It also had the classic structural trait of SOCS family,in which a central SH2 domain,a C-terminal 40-amino-acid SOCS box domain and a 12-amino-acid KIR domain were consisted.The dynamic of SOCS3 mRNA in thymus gland,spleen,mesenteric lymph node and intestine lamina propria of pigs during 10 days consecutive heat stress was investigated.The expression of SOCS3 in mesenteric lymph node were decreased on day 3,6and 9after heat stress begin significantly(P≤0.05),but a drastic up-regulation on day 6and 9in the spleen and intestine lamina propria,although a reduced tendency was displayed on day 3(P≤0.05).The porcine SOCS3 cDNA was cloned successfully,and a significant changes of SOCS3 mRNA expression was investigated in heat stressed pigs with a tissue and time dependence manner.