猪-人移植细胞性排斥反应相关候选基因pOSR1的克隆和鉴定

为寻找与异种移植细胞性排斥反应相关的新基因 ,在抑制消减杂交 (SSH )基础上 ,对一个与人氧化应激反应 1(OSR1)基因同源的猪内皮细胞上调表达基因片段CS3作进一步研究。半定量RT PCR结果证实 ,在人PBMC作用后 ,该基因表达的确上调。应用快速cDNA末端扩增 (RACE )方法 ,获得了猪OSR1(pOSR1)的全长cDNA序列。pOSR1cDNA序列全长 4 333个碱基 ,开读框架长 15 90个碱基 ,编码 5 2 9个氨基酸。与人OSR1(hOSR1)相比 ,编码区核酸一致性为 92 8% ,氨基酸一致性为 95 5 %。GenBank登录号为AY2 7135 6。其功能研究正在进行中。

Cloning and Identification of the Candidate Gene pOSR1 Associated with Cell-Mediated Pig to Human Xenotransplantation

In order to find out the novel gene associated with the cell-mediated pig to human xenograft rejection,gene fragment CS3 associated with the up-regulated expression of the porcine endothelial cells and homologous with the human oxidative stress responsive 1 (OSR1) was further investigated on the basis of suppression subtractive hybridization (SSH). Results of semi-quantitative RT-PCR demonstrated that this gene was really up-regulated in the interactions of human PBMC and porcine endothelial cells. The full-length cDNA sequence of porcine OSR1 (pOSR1) with a length of 4 333 bp was obtained through the rapid amplification of cDNA ends (RACE) technique,and the open reading frame (ORF) with a length of 1 590 bp encoded 529 amino acid residues was also obtained. This gene showed 92.8% of nucleotide identity and 95.5% of amino acid identity with human OSR1. The GenBank accession number of this gene is AY271356 and the study on the function of this gene is still undergoing.