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2025年8月13日 星期三
  2017, Vol. 25 Issue (5): 770-780    
  研究论文与报告 本期目录 | 过刊浏览 | 高级检索 |
实验用SPF大白猪和长白猪SLA-1基因特征分析
权金强1,江新杰2,李昌文2,路小野2,陈洪岩2,赵生国1,高彩霞2
1. 甘肃农业大学
2.
Characterization Analysis of SLA-1 Gene from SPF Yorkshire (Sus scrofa) and Landrace
1, 1, 1, 1, 1, 2, 1
1.
2. Gansu Agricultural University
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摘要 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cDNA,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。
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权金强
江新杰
李昌文
路小野
陈洪岩
赵生国
高彩霞
关键词 无特定病原体(SPF)大白猪SPF长白猪猪白细胞抗原(SLA)基因特征    
Abstract:Abstract Swine (Sus scrofa) leukocyte antigen (SLA) gene is one of the genetic factors that cause xenotransplantation cell immune rejection. To enrich the classical experimental animal model and solve the problem of the lack of heterotopic transplantation model, blood samples were obtained from specific pathogen free (SPF) Yorkshire and Landrace for isolation of peripheral blood mononuclear cells, and SLA-1 gene was amplified by PCR after reverse transcription. SLA-1 gene sequences were obtained by analyzing the direct and cloning results. The polymorphism of alleles was analyzed using the bioinformatics software. A total of 16 alleles was identified at SLA-1 gene, containing a full-length 1 086 bp open reading frame. Nucleotide diversity (Pi) was 0.045 59, haplotype diversity (Hd) was 0.000 49, the average nucleotide difference (K) was 49.508, and the content of G+C reached 60.9%. Among 150 nucleotide mutations, the number of nucleotide parsimony informative sites (NPi) were higher than those of the nucleotide singleton variable sites (NS), and 82 of the 361 amino acids were mutated and the numbers of non-synonymous sites (NSS) were obviously higher than those of synonymous sites (SS). The polymorphism of SLA-1 allele was mainly concentrated in exon 2 and 3. Nine of 33 key amino acids were completely conserved, and eight residues showed complete consistence with the corresponding amino acids of human leukocyte antigen (HLA) allele. Eleven of the 19 amino acids, which bind β2-microglobulin (β2m) to HLA-A, showed complete consistency among the alleles identified in this study. The key amino acid binding sites of CD8 to major histocompatibility complex (MHC) were highly consistent in human (Homo sapiens) and SPF Yorkshire and Landrace, except for 225 (Thr/Ser) and 228 (Thr/Met) sites. The result of phylogenetic relationship showed that there was a close relationship among SPF Yorkshire and Landrace pigs, Korean native pigs and Meishan pigs. This study can provide genetic basis for establishment of animal models antigen presentation and immune response.
Key wordsSpecific pathogen free (SPF) Yorkshire    SPF Landrace    Swine leukocyte antigen (SLA) gene    Characterization
收稿日期: 2016-12-12      出版日期: 2017-05-02
基金资助:国家自然科学基金
通讯作者: 赵生国     E-mail: zhaosg@gsau.edu.cn
引用本文:   
权金强 江新杰 李昌文 路小野 陈洪岩 赵生国 高彩霞. 实验用SPF大白猪和长白猪SLA-1基因特征分析[J]. , 2017, 25(5): 770-780.
链接本文:  
http://journal05.magtech.org.cn/Jwk_ny/CN/     或     http://journal05.magtech.org.cn/Jwk_ny/CN/Y2017/V25/I5/770
 
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