苏紫猪SLA-1基因多态性及其抗病潜能分析

doi:10.16431/j.cnki.1671-7236.2023.05.019

摘要/Abstract

摘要： 【目的】探究苏紫猪白细胞抗原-1(swine leukocyte antigen-1,SLA-1)基因多态性,并对其抗病潜能进行分析,以期为苏紫猪分子育种提供理论基础。【方法】以苏紫猪血细胞cDNA为模板,用PCR法扩增苏紫猪SLA-1基因并测序,利用DNAStar和Mega 5.0软件分别进行相似性分析及系统进化树构建;利用NetMHCpan 4.1 server分析SLA-1与非洲猪瘟病毒(African swine fever virus,ASFV)抗原表位结合的能力及特征;运用生物学软件对其编码蛋白的理化性质、亲/疏水性、跨膜区、结构和功能等进行预测。【结果】试验成功扩增6条苏紫猪SLA-1等位基因序列,分别为SLA-1*sz01、SLA-1*sz02、SLA-1*sz03、SLA-1*sz04、SLA-1*sz05和SLA-1*sz06,其高变异位点主要位于肽结合沟(peptide-binding groove,PBG)α1区和α2区;相似性比对结果显示,6条苏紫猪SLA-1等位基因核苷酸、氨基酸序列相似性分别在92.9%~96.9%、87.0%~94.8%之间,与不同品种猪核苷酸、氨基酸序列相似性分别在92.8%~98.8%、84.3%~98.0%之间;ASFV来源多肽结合能力的预测结果显示,SLA-1*sz01蛋白具有较强的ASFV抗原呈递能力,与ASFV来源B354L蛋白多肽片段FIDKTTVLY具有最高亲和力;SLA-1*sz01蛋白功能域及三级结构预测结果显示,SLA-1*sz01与免疫系统相关,具有经典的SLA-1三级结构。【结论】本研究扩增得到6条苏紫猪SLA-1基因序列,其中SLA-1*sz01能够限制性结合较多的ASFV来源抗原表位多肽,与多肽片段FIDKTTVLY具有最高亲和力,可作为苏紫猪抗病育种的候选基因。研究结果为ASFV疫苗的研发提供参考基础。

关键词: 苏紫猪; SLA-1基因; 多态性; 抗原结合位点; 抗病潜能

Abstract: 【Objective】 The purpose of this study was to explore the genetic polymorphism of swine leukocyte antigen-1(SLA-1) and analyze its disease resistance potential,in order to provide a theoretical basis for molecular breeding of Suzi pig.【Method】 The SLA-1 gene was amplified by PCR using blood cell cDNA in Suzi pig as a template and sequenced. Similarity analysis and phylogenetic tree construction were conducted using DNAStar and Mega 5.0 software, respectively.NetMHCpan 4.1 server was used to analyze the ability and characteristics of SLA-1 molecule binding to African swine fever virus (ASFV) antigenic epitopes.The physical and chemical properties,hydrophilic and hydrophobic properties,transmembrane region,structure and function of the encoded proteins were predicted by biological software.【Result】 Six SLA-1 allele sequences of Suzi pig were successfully amplified,and namely SLA-1*sz01,SLA-1*sz02,SLA-1*sz03,SLA-1*sz04,SLA-1*sz05 and SLA-1*sz06.The highly variable sites were mainly located in the α1 and α2 regions of the peptide binding groove (PBG).The similarity comparison results showed that the similarity of the six SLA-1 alleles nucleotide and amino acid sequences of Suzi pig were between 92.9% to 96.9% and 87.0% to 94.8%,respectively.The similarity of the six SLA-1 alleles nucleotide and amino acid sequences of Suzi pig with different pig breeds were between 92.8% to 98.8% and 84.3% to 98.0%,respectively.The prediction results of the binding ability of ASFV derived polypeptides showed that the SLA-1*sz01 protein had a strong ASFV antigen presentation ability,and had highly affinity with B354L protein polypeptide fragment FIDKTTVLY in ASFV.Prediction of functional domain and tertiary structure showed that SLA-1*sz01 was related to the immune system and had a classic SLA-1 tertiary structure.【Conclusion】 In this study,six SLA-1 gene sequences were obtained,among which SLA-1*sz01 could restrict the binding of more ASFV-derived peptides,and had the highest affinity with FIDKTTVLY,which could be used as a candidate gene for disease resistance breeding of Suzi pig.The results provided a reference basis for the research and development of ASFV vaccine.

Key words: Suzi pig; SLA-1 gene; polymorphism; antigen binding site; disease resistance potential

中图分类号:

S813.3

王泽平, 王飞飞, 张芮铭, 付言峰, 赵为民, 戴超辉, 程金花, 廖超, 李辉. 苏紫猪SLA-1基因多态性及其抗病潜能分析[J]. 中国畜牧兽医, 2023, 50(5): 1918-1927.

WANG Zeping, WANG Feifei, ZHANG Ruiming, FU Yanfeng, ZHAO Weimin, DAI Chaohui, CHENG Jinhua, LIAO Chao, LI Hui. Analysis on Polymorphism of SLA-1 Gene and Its Disease Resistance Potential in Suzi Pig[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(5): 1918-1927.

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