实验用SPF大白猪和长白猪SLA-1基因特征分析

发布时间：2018-05-26 13:05

本文选题：无特定病原体(SPF)大白猪 + SPF长白猪　；参考：《农业生物技术学报》2017年05期

【摘要】：猪(Sus scrofa)白细胞抗原(swine leukocyte antigen,SLA)基因是引起异种移植细胞性免疫排斥的遗传因素之一。为丰富经典实验动物模型,解决异种器官移植模型缺乏问题,本研究采取实验用无特定病原体(specific pathogen free,SPF)大白猪和长白猪抗凝血,分离外周血淋巴细胞,提取总RNA,反转录c DNA,设计特异性引物进行PCR扩增测序,获得SLA-1基因,并进行特征分析。本研究共鉴定了16个SLA-1等位基因,各含有一个全长1 086 bp的完整开放阅读框,核苷酸多样度(nucleotide diversity,Pi)为0.045 59,单倍型多样度(haplotype diversity,Hd)为0.000 49,平均核苷酸差异数(average number of nucleotide differences,K)为49.508,G+C含量高达60.9%。150个核苷酸变异位点中简约信息位点(nucleotide parsimony informative sites,NPi)远高于单一多态位点(nucleotide singleton variable sites,NS);共编码361个氨基酸,产生82个突变,非同义突变(non-synonymous mutation sites,NSS)远高于同义突变(synonymous mutation sites,SS)。SLA-1等位基因多态性集中在第2和3外显子,33个关键氨基酸有9个保守位点,与人白细胞抗原-A(human leukocyte antigen-A,HLA-A)基因相同的位点有8个。HLA-A与β2-微球蛋白(β2-microglobulin,β2m)结合的19个氨基酸中,11个氨基酸与SPF大白猪和长白猪保持一致;人(Homo sapiens)和SPF大白猪、长白猪CD8分子与主要组织相容性复合体(major histocompatibility complex,MHC)结合的关键氨基酸位点高度保守,只有225(Thr/Ser)和228(Thr/Met)位点不同。系统进化树表明,SPF大白猪和长白猪与Yucatan、韩国本地猪和梅山猪亲缘关系较近。本研究成功鉴定了16个SLA-1等位基因,高度多态,且SLA保留了HLA的部分功能位点。研究结果为构建动物模型以及相关的抗原递呈和免疫应答反应提供遗传学参考依据。
[Abstract]:Sus scrofaa (Sus scrofaa) WBC antigenine leukocyte antigeno gene is one of the genetic factors that cause cellular immune rejection in xenografts. In order to enrich the classical experimental animal model and solve the problem of lack of xenogeneic organ transplantation model, in this study, anticoagulant blood was isolated from peripheral blood lymphocytes of large white pigs and Landrace pigs without specific pathogen. Total RNAs were extracted, reverse transcripted c-DNAs were extracted, specific primers were designed for PCR amplification and sequencing, SLA-1 gene was obtained, and characteristic analysis was carried out. In this study, 16 SLA-1 alleles were identified, each containing a complete open reading frame with a length of 1 086 BP. The nucleotide diversity of nucleotide was 0.045 59, the haplotype site was 0.000 49, and the average nucleotide difference number was 49.508 G / C, which was much higher than that of nucleotide parsimony informative sitesophane (NPI). Nucleotide singleton variable sitesngs, encoding 361 amino acids, The allele polymorphism of non-synonymous mutation sites-1 was significantly higher than that of synonymous mutation SLA-1 in exons 2 and 3, and there were 9 conserved sites in 33 key amino acids. Among the 19 amino acids of HLA-A binding to 尾 2-microglobulin (尾 2 m), 11 amino acids were identical with SPF white pigs and Landrace pigs, and human homo sapiens and SPF white pigs. The key amino acid sites for the binding of Landrace pig CD8 molecule to major histocompatibility complex of major histocompatibility complex (MHC) are highly conserved, with only 225 THR / Sera and 228 THR / Metals different sites. Phylogenetic tree showed that SPF and Landrace pigs were closely related to Yucatan, Korean native pigs and Meishan pigs. In this study, 16 SLA-1 alleles were identified with high polymorphism, and SLA retained some functional sites of HLA. The results provide genetic reference for the establishment of animal models and related antigen presentation and immune response.
【作者单位】：甘肃农业大学动物科学技术学院;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;牡丹江师范学院生命科学与技术学院;
【基金】：国家自然科学基金(No.31502039)
【分类号】：S828

【相似文献】