Molecular cloning and characterization of porcine indoleamine 2, 3-dioxygenase and its expression in various tissues
Molecular cloning and characterization of porcine indoleamine 2, 3-dioxygenase and its expression in various tissues
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猪吲哚胺2,3-双加氧酶的分子克隆、表征及其在各组织中的表达
DOI：
10.1007/s11596-012-0082-y
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发表时间：
2012-08-01
影响因子：
--
通讯作者：
Zhu, Min
中科院分区：
文献类型：
--
作者：
Chen, Chao;Wei, Mingfa;Zhu, Min
为了证实猪中存在吲哚胺2，3-双加氧酶（IDO）基因，并克隆该新基因，分析其分子结构特征和表达模式，利用生物信息学方法从GenBank数据库中获得了猪IDO基因的同源序列。利用RT-PCR技术从猪血管内皮细胞系中克隆了IDO基因，并对其进行了序列测定和表达谱分析。参考人IDO的三级结构，利用生物序列分析软件和数据库建立猪IDO的三维结构模型。结果表明，猪IDO基因经测序鉴定正确。该基因序列经Genbank登录后证实为一个新基因。猪IDO基因在肺、胸腺、附睾和前房中有基础表达，在外周血单个核细胞中有高表达。猪IDO的三维结构模型与人IDO的三维结构模型相似。因此，鉴定猪IDO基因的结构信息，对进一步研究该基因的免疫功能具有重要意义。IDO对NK细胞介导的异种移植排斥反应的研究将为异种移植的发展提供新的治疗靶点。
In order to confirm the existence of indoleamine 2, 3-dioxygenase (IDO) gene in swine, and to clone the novel gene followed by the molecule structure properties and expression pattern analysis, the porcine mRNA sequences homologous to human IDO were obtained from GenBank database by bioinformatics method. By using RT-PCR, the IDO gene was cloned from porcine endothelial cell line and the accuracy of the nucleic acid sequence was confirmed, and the expression pattern of the gene was detected. The three-dimensional structure model of porcine IDO was built referring to the tertiary structure of human IDO using biological sequence analysis software and database. The results showed that the porcine IDO was identified by sequencing. The nucleotide sequences were confirmed as a novel gene after submitted to Genbank. Porcine IDO was expressed in the lung, thymus, epididymis and anterior chamber with a basic level, however in peripheral blood mononuclear cells (PBMCs) the IDO gene was highly expressed. The three-dimensional structure model of porcine IDO was similar to that of human IDO. It was suggested that identification of the structure information of porcine IDO is essential to further investigate the immunologic function of the gene. Study of IDO on NK cells-mediated xenograft rejection will be a novel therapeutic target for the development of xenotransplantation.