作　　者： (江新杰）; (权金强）; (李昌文）; (路小野）; (陈洪岩）; (杨春文）; (高彩霞）;

机构地区： 牡丹江师范学院生命科学与技术学院,黑龙江省牡丹江157012 中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室,哈尔滨150069

出　　处： 《中国实验动物学报》 2017年第4期404-413,共10页

摘　　要： 目的研究实验用SPF大白猪和长白猪SLA II类基因的多态性。方法分别采集15头SPF大白猪和22头长白猪抗凝血,分离外周血淋巴细胞,提取总RNA,反转录后RT-PCR扩增DQB1、DRB1和DQA基因并进行测序,分析获得的SLA II类等位基因序列多态性。结果 3个基因共获得25个等位基因,包括8个DQB1,10个DRB1和7个DQA,全部获得ISAG SLA命名委员会的官方命名,其中3个等位基因首次提交完整序列,命名为DQB1*02:12(KU754590)、DQB1*02:03(KU754591)和DRB1*06:07(KU754601),3个DQA等位基因为新发现等位基因。SPF大白猪和长白猪DQB1等位基因与外源性抗原结合的15个氨基酸中,共有5个氨基酸具有高度保守性;DRB1等位基因的16个外源性抗原识别位点中,仅1个位点高度保守;DQA等位基因19个抗原结合位点中,有11个高度保守。SLA II类基因氨基酸序列分子进化树表明,3个基因分别聚为两大类,与国外Yucatan小型猪具有较近的亲缘关系,而与其他猪种未表现明显遗传距离相隔。结论成功鉴定了大白猪和长白猪的25个SLA II类等位基因,发现其具有较为丰富的多态性,所获得SLA II类等位基因在其他品种猪也广泛分布,具有多样性,这一研究结果对大白猪和长白猪发展为经典实验动物模型具有重要作用。 Objective The aim of this study was to investigate the polymorphism of SLA class II genes in Canadian SPF Yorkshire and Landrace pigs. Methods Blood samples were obtained from 15 SPF Yorkshire and 22 Landrace pigs for isolation of peripheral blood mononuclear cells respectively,and the DQB1,DRB1 and DQA genes were amplified by PCR after reverse transcription. SLA class II genes were obtained by analyzing the direct and cloning result. The polymorphism of alleles was analyzed using the DNAsp 5. 0 software. Results A total of 25 alleles were identified at threegenes,including eight DQB1,ten DRB1 and seven DQA,and three alleles were submitted the complete sequences for the first time. The official allele names were assigned as SLA-DQB1 ＊ 0212（ KU754590）,SLA-DQB1 ＊ 0203（ KU754591）and DRB1＊ 06： 07（ KU754601） by the SLA Nomenclature Committee. Three novel DQA alleles were discovered. Five of the 15 amino acids,one of the 16 amino acids and 11 of the 19 amino acids,which bind processing antigens,showed well conserved among the alleles of DQB1,DRB1 and DQA genes in the SPF Yorkshire and Landrace pigs,respectively. Neighbor-joining tree showed that the three genes were divided into two clusters,respectively. There was a close relationship between SPF Yorkshire and Landrace pigs and foreign Yucatan miniature pigs,and it showed no obvious genetic distance with other pigs. Conclusions A total of 25 SLA class II alleles have been identified successfully in this study,and there are more abundant polymorphism for them. There is a widely distribution for SLA class II alleles identified in this study in other pig breeds. It is critical for the eventual future use of SPF Yorkshire and Landrace pigs as classical laboratory animal models.

关 键 词： 猪白细胞抗原 大白猪 长白猪 类等位基因 多态性