广西巴马小型猪白细胞抗原经典Ⅱ类基因克隆及生物信息学分析

摘要

为探讨广西巴马小型猪SLA经典Ⅱ类基因分子结构特征及其在猪-人异种器官移植研究中的应用前景,本研究采集4头广西巴马小型猪的耳组织,经抽提RNA和反转录得到cDNA,通过PCR扩增,对SLA经典Ⅱ类基因( SLA- DRA、SLA- DRB1、SLA-DQA、SLA-DQB1)纯化和克隆,进行双向测序.结果,在4个SLA经典Ⅱ类座位中得到基因序列共8条,利用NCBI服务器中BLAST分析发现,8条基因序列中5条为已知的等位基因,另3条为新的等位基因.通过核苷酸和氨基酸水平的同源性比对,构建亲缘进化树,分析发现,广西巴马小型猪具有与人同源性较高的MHC遗传结构,显示该猪种在人-猪异种移植领域中有着潜在的利用价值.%The study aimed to elucidate the structures of SLA Class ? genes of Guangxi Bama Minipigs and the potential in pigs to humans xenotransplantation. 4 specimens of ear tissues were sampled from four Guangxi Bama Minipigs, and the classical swine leukocyte antigens class ? genes (SLA-DRA, SLA-DRB1, SLA-DQA , SLA-DQB1) were amplified by PCR after reverse transcription. Purified products were cloned into pMD18-T vectors and bi-directional sequenced. Analyzing obtained sequences by BLAST in NCBI, a total of eight alleles were found, and five alleles have been found in previous reports, the other three were novel. From the homology analysis between alleles of SLA and HLA in GenBank and phylogenetic trees analysis, the results showed that the Guangxi Bama Minipigs possessed high homologous in MHC information with human, and indicated that Guangxi Bama minipigs would have potential utilization values in the study of pig to human xenotransplantation.