广西巴马小型猪LAIR-1基因cDNA克隆及序列分析

Cloning and sequence analysis of cDNA of leukocyte-associated immunoglobulin-like receptor-1 (LAIR-1) gene in Guangxi Bama mini-pig

【目的】了解广西巴马小型猪白细胞相关免疫球蛋白样受体1(LAIR-1)基因的序列特征及基本结构,为以广西巴马小型猪作为人类器官模型及器官移植手术后的排斥反应研究奠定基础。【方法】根据GenBank上已登录的人和家鼠LAIR-1基因序列的保守区域,用Oligo设计合成1对引物,并以广西巴马小型猪总RNA为模板,对LAIR-1基因进行RT-PCR扩增,经PCR和双酶切鉴定后,进行测序及同源性比对分析。【结果】从广西巴马小型猪的外周血淋巴细胞中克隆获得LAIR-1基因cDNA序列,全长867bp;与人、家鼠和挪威鼠的同源性分别为79.3%、50.9%和49.9%。【结论】广西巴马小型猪LAIR-1基因cDNA序列与人类的亲缘关系最近,进一步验证了广西巴马小型猪作为人类器官模型研究的可行性。

[Objective ]Sequence characteristics and basic structure of leukocyte-associated immunoglobulin -like receptor （LAIR-1 ） were analyzed to lay the foundation for research on rejection response after organ transplantation of Guangxi Bama min-pig by using them as human organ model. [Method]In this experiment, according to the conservative region of LAIR-lgene sequence in the human and mouse registered in GeneBank, a pair of primers was designed and composed by Oligo. LAIR-I gene was amplified using RT-PCR using total RNA template from GuangxiBama mini-pig. Sequencing and homology analyses were performed after PCR sequencing and double digestion identification. [ Result]The overall length of cDNA sequence of LAIR-1 gene, cloned from peripheral blood lymphocytes of Bama mini-pig, was 867 bp,which shared 79.3, 50.9 and 49.9% of homology with human, house mouse and Norwegian mouse, respectively. [Conclusion]Due to its closest genetic relationship with human, Guangxi Bama mini-pig was confirmed feasible to be used as human organ model in research.