短尾蝮蛇IRF-2基因的克隆和特征分析

Cloning and Character Analyses of IRF-2 Gene for Gloydius brevicaudus

报道了短尾蝮蛇Gloydius brevicaudus干扰素调节因子2基因(IRF-2)的克隆和cDNA全序列测定分析。目前除在爬行类外,IRF-2基因在大部分的脊椎动物如鱼类、两栖类、鸟类和哺乳类都有报道。为了获得爬行类IRF-2基因的全序列,从短尾蝮蛇的肾、脾、肝和肠4种组织中使用Trizol试剂盒提取了总RNA。从已知的IRF-2基因序列比对设计了简并引物来扩增保守片段。最后使用RACE方法得到IRF-2的cDNA全序列。结果显示短尾蝮蛇的IRF-2基因开放阅读框包含有978个核苷酸,编码326个氨基酸,其3′UTR包含137个核苷酸。与脊椎动物其他四足动物序列比对分析还发现,短尾蝮蛇的IRF-2基因和推导的氨基酸序列都非常保守,与大部分四足动物的IRF-2相似。从其氨基酸序列中可以辨别出DBD、IDA2和transactivating dom ain等大部分元件和阻遏模体。

IRF-2 gene have been reported in most of vertebrates such as fishes,amphibians,birds and mammals but not in reptiles.For the purpose of obtaining the cDNA sequence of reptile IRF-2,total RNA was isolated from the tissues of kidney,spleen,liver,and intestine of a viper（Gloydius brevicaudus） using Trizol kit.The degenerate primers to obtain IRF-2 were designed by comparing known IRF-2 sequences.RACE method also was applied to obtain the complete cDNA sequence of IRF-2 gene.The complete cDNA sequence of G.brevicaudus IRF-2 gene contains an ORF of 978 nt which encoding 326 amino acid with a 3＇ UTR of 137 nt.Aligned with other IRF-2 gene sequence,we found that reptiles have very similar sequence of IRF-2 with other vertebrates,especially the first 345 nt of ORF are very conserved due to they are DNA banding domain.And some elements,such as DBD,IAD2,transactivating domain and repression motif were identified in the amino acid sequence of G.brevicaudus IRF-2.