猪SOCS4基因的克隆、生物信息学分析及热应激条件下的表达

Cloning and Bioinformatics Analysis of Porcine SOCS4 cDNA and Its Dynamic during Heat Stress

旨在研究和系统阐述热应激诱发猪免疫抑制及炎性因子紊乱的发生机理。本研究克隆了土杂(陆川猪×长白猪)猪SOCS4基因,发现其开放阅读框(ORF)长1 326bp,编码441个氨基酸,N端包含285个氨基酸,与长白猪、牛、山羊、猫、马、犬、人和小鼠的氨基酸序列同源性分别为100.0%、96.1%、96.1%、95.2%、94.6%、94.3%、93.0%和88.1%;蛋白分子量为50.5ku,等电点(PI)为6.60。荧光定量PCR结果显示,高温条件下,第3和第9天胸腺组织上SOCS4 mRNA显著上调表达(P≤0.05),而在第1和第6天的肠系膜淋巴结中极显著上调表达(P≤0.01)。与上述组织不同,SOCS4mRNA在小肠和脾组织中却显著下调表达(P≤0.05)。本研究成功克隆了土杂猪SOCS4基因,发现其在高温条件下的表达量发生明显改变,并呈组织和时间依赖性。

The aim of this study was to systematically elucidate the mechanism of heat stress-induced swine immunosuppression,and inflammatory factors disorder.The TZ(Luchuan pig×Landrace pig)porcine SOCS4 gene was cloned in this study,it consisted of 1 326 bp of ORF encoding aprotein with 441 amino acids,285 amino acids of them formed the N-terminal of SOCS4.It shared high amino acid sequence identity with that in Landrace pigs(100.0%),Bos Taurus(96.1%),Capra Hircus(96.1%),Felis catus(95.2%),Equus caballus(94.6%),Canis lupus familiaris(94.3%),Homo sapiens(93.0%)and Mus musculus(88.1%).The molecular weight of SOCS4 protein was 50.5ku,and PI was 6.60.qRT-PCR results showed that the expression of SOCS4 in thymus gland of TZ pigs was significantly upregulated on day 3and 9after heat stressbegan(P≤0.05).As well in mesenteric lymph node,where it significantly upregulated on day 1and 6(P≤0.01).The expression of SOCS4 decreased significantly in small intestine and spleen of TZ pigs during heat stress(P≤0.05).The TZ porcine SOCS4 cDNA was cloned successfully,and a significant changes of SOCS4 mRNA expression were appeared in heat stressed pigs with a tissue and time dependence manner.