[目的]通过生物信息学方法分析1株2022年辽宁沈阳地区猫泛白细胞减少症病毒(Feline panleukopenia virus,FPV)LN3株VP2蛋白的分子特征.[方法]利用FPV胶体金试纸条对临床上表现呕吐、腹泻等症状的患病猫粪便进行检测,提取该病猫粪便的病毒DNA进行FPV VP2基因PCR扩增及测序,使用Seqman软件对序列进行拼接.将所获序列与NCBI数据库中FPV和犬细小病毒(Canine parvovirus,CPV)的VP2基因进行相似性比对及遗传进化分析.使用生物信息学软件对该毒株VP2蛋白进行预测,包括理化性质、亲/疏水性、跨膜区、糖基化位点、亚细胞定位、二级结构、三级结构等.[结果]FPV LN3株VP2基因全长1 755 bp,编码584个氨基酸;与GenBank上登录的15株FPV同属一个大分支,相似性为98.9％～99.7％;与CPV处于不同分支上.生物信息学软件预测显示,FPV LN3株VP2蛋白为亲水性蛋白,无跨膜结构;含有7个潜在N-糖基化位点、86个O-糖基化位点和50个磷酸化位点;VP2蛋白在细胞质、细胞核、线粒体中的可能性分别为43.5％、34.8％和21.7％;VP2蛋白二级结构中无规则卷曲、a-螺旋、延伸链、β-转角分别占61.82％、8.90％、24.32％及4.97％,三级结构预测结果与其一致;VP2蛋白共有20个抗原表位.[结论]VP2是FPV遗传变异的关键基因,FPV LN3株VP2蛋白属于亲水性、非跨膜稳定蛋白,共存在20个抗原表位.试验结果为进一步研究FPV VP2蛋白功能及研发新型疫苗提供理论依据.
[Objective]The purpose of this study was to analyze the molecular characteristics of VP2 protein of a Feline panleukopenia virus(FPV)LN3 strain from Shenyang,Liaoning province in 2022 by bioinformatics methods.[Method]FPV colloidal gold test strips were used to detect feces of sick cats with clinical symptoms such as vomiting and diarrhea.The extracted viral DNA from the feces of the diseased cat was amplificated by PCR and sequenced.Sequences were spliced by Seqman software.The VP2 sequence of FPV LN3 strain was subjected to similarity alignment and genetic evolution analysis with FPV and Canine parvovirus(CPV)strains in NCBI.The VP2 protein of FPV LN3 strain was predicted by bioinformatics software,included physical and chemical properties,hydrophilicity and hydrophobicity,transmembrane region,glycosylation site,subcellular localization,protein secondary structure,tertiary structure,etc.[Result]The full length of VP2 gene of FPV LN3 strain was 1 755 bp,which was composed by 584 amino acids.The FPV LN3 strain and other 15 strains registered on GenBank belong to the same major branch,with similarity ranging from 98.9％to 99.7％.FPV LN3 strain and CPV were not belong to the same branch.Bioinformatics software prediction results showed that the VP2 protein of FPV LN3 strain was hydrophilic and had no transmembrane structure.VP2 protein contained 7 potential N-glycosylation sites,86 O-glycosylation sites and 50 phosphorylation sites.The probability of VP2 protein in cytoplasm,nucleus and mitochondria were 43.5％,34.8％and 21.7％,respectively.The randon coil,alpha helix,extended chain and beta turn in the secondary structure of VP2 protein accounted for 61.82％,8.90％,24.32％and 4.97％,respectively,which was consistent with the prediction results of the tertiary structure.There were 20 antigenic epitopes of VP2 protein.[Conclusion]VP 2 was a key gene for genetic variation in FPV.The VP2 protein of FPV LN3 strain was a hydrophilic and non transmembrane stable protein,which contained 20 antigenic epitopes.The results could provide theoretical basis for further study of the function of FPV VP2 protein and the development of new vaccines.
刘琪;刘正伟;梁琳;张利;郝春晖;李玥;赵福庆
辽宁农业职业技术学院,营口 115009中国农业科学院北京畜牧兽医研究所,北京 100193
畜牧业
猫泛白细胞减少症病毒;VP2基因;生物信息学;蛋白结构
Feline panleukopenia virus(FPV);VP2 gene;bioinformatics;protein structure
《中国畜牧兽医》 2024 (001)
23-32 / 10
辽宁省自然科学基金项目-营口联合基金(2021-YKLH-11);辽宁农业职业技术学院2021年院级科研项目(Lnz50)
10.16431/j.cnki.1671-7236.2024.01.003
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