摘要
【目的】研究猪流行性腹泻病毒(porcine epidemic diarrhea virus,PEDV)在抗体选择压下的分子变异。【方法】将PEDV Sinder02株在添加适量抗体和不添加抗体的Vero E6细胞上连续传代40代,对传代前后毒株的全基因组序列进行扩增、测序、拼接和分析。【结果】第40代有抗体组和无抗体组全基因组核苷酸发生突变的位点累计分别为71和50个,氨基酸发生突变的位点累计分别为50和26个。有抗体组S基因的非同义突变(nonsynonymous mutation,NS)与同义突变(synonymous mutation,S)的比值(NS/S值)为4.25,ORF3基因的NS/S值为3.67。有抗体组S基因出现6个稳定突变位点,其中4个与已知抗原表位有关;ORF3基因出现5个稳定突变位点,其中3个与已知抗原表位有关。有抗体组、无抗体组与原代病毒序列的同源性逐代降低,且有抗体组与原代病毒之间的差距更大、遗传距离更远。【结论】首次证明抗体选择压对PEDV Sinder02株的S基因和ORF3基因起到促变异作用。研究结果为探索PEDV变异提供了理论基础,也为新型PEDV疫苗的研制提供了科学参考。
[Purpose]To study the molecular variation of porcine epidemic diarrhea virus(PEDV)under antibody selective pressure.[Methods]PEDV Sinder02 strain was continuously passaged for 40 generations on Vero E6 cells with and without the addition of appropriate antibodies,and the whole genome sequences of the virulent strains before and after passaging were amplified,sequenced,spliced,and analyzed.[Results]In the 40th generation,the cumulative number of genome nucleotide mutation sites respectively was 71 and 50 for the group with antibody(group A)and the group without antibody(group NA),and the cumulative number of amino acid mutation sites respectively was 50 and 26 for the group A and the group NA.The ratio of nonsynonymous(NS)to synonymous(S)in S gene of the group A was 4.25,and the NS/S value in ORF3 gene was 3.67.In the group A,there were six stable mutation sites in S gene,four of which were associated with known epitopes;and there were five stable mutation sites in ORF3 gene,three of which were associated with known epitopes.The sequence homology between the original virus and both the group A and the group NA were decreased from generation to generation,and the differences between the group A and the original virus was greater,with a greater genetic distance.[Conclusion]The first proof that antibody selective pressure plays a role in promoting variation of the S gene and ORF3 gene of PEDV Sinder02 strain.The results of the study provide a theoretical basis for exploring the variation of PEDV and also provide a scientific reference for the development of new PEDV vaccines.
作者
余蕊
高艺
孙永科
张恒
杨玉艾
YU Rui;GAO Yi;SUN Yongke;ZHANG Heng;YANG Yuai(College of Veterinary Medicine,Yunnan Agricultural University,Kunming 650201,China;Pig Disease Research Center,Shandong Sinder Technology Co.,Ltd.,Qingdao 266104,China)
出处
《云南农业大学学报(自然科学版)》
北大核心
2025年第1期79-85,共7页
Journal of Yunnan Agricultural University:Natural Science
基金
国家自然科学基金项目(32160845)。
