猪SOCS-2基因的克隆及序列分析

从中国地方猪品种八眉猪(BaMei)肾脏组织中提取总RNA,采用RT-PCR方法克隆了猪SOCS-2(suppressor of cytokinesignaling-2,细胞因子信号转导抑制因子-2)基因的cDNA序列,经T/A克隆,插入到pMD19-T载体上,导入大肠杆菌DH-5α,阳性克隆经PCR鉴定后进行测序,将测序结果与GenBank中已登录的人、大鼠和小鼠SOCS-2基因的序列进行同源性比较,利用生物信息学和分子生物学软件对猪SOCS-2基因编码的蛋白进行结构预测。结果表明:首次成功克隆了猪SOCS-2基因的cDNA序列(GenBank登录号为EF121242),其长度为822bp,该基因ORF区核苷酸序列与其他物种相比同源性达到93%以上,氨基酸同源性则达到89%以上,生物信息学分析表明该蛋白分子量为22.25kD,等电点pI=8.30,包含199个氨基酸残基。该基因cDNA序列的克隆,有利于进一步研究SOCS-2调节机体发育的分子机理。

Cloning and Sequence Analysis of SOCS-2 Gene in Pig

Total RNA was isolated from kidney of BaMei pig, a local strain of Chinese pig, and then the cDNA sequence of SOCS-2 gene was cloned by RT-PCR (GenBank accepted number is EF121242). Then the cloned SOCS-2 gene was inserted into PMD19-T vector by T/A cloning, transformed into DH-5alpha, tested by PCR and sequenced. The data show that the homology of the cloned porcine SOCS-2, including 822 bp, is more than 93% and that of the deduced amino acid sequence is 89% when compared with human, rat and mice. And the molecular weight of SOCS-2 protein is about 22.25 kD and PI is 8.03. The cloning of SOCS-2 gene is useful for the further research on the molecular mechanism by which regulating growth and development of organism.