摘要
为阐述藏猪猪细小病毒(PPV)非结构蛋白1(NS1)和结构蛋白2(VP2)的生物信息学特征,本试验对疑似患病西藏本土藏猪血液样本进行聚合酶链式反应(PCR)和全基因组测序鉴定,阳性样本进行病毒分离并观察细胞病变效应(CPE),绘制PPV分离株遗传进化树进行亲缘性分析;运用在线网站和软件对PPV分离株的NS1和VP2蛋白氨基酸进行生物信息学分析,获取2种蛋白的理化性质和空间结构。结果显示,经PCR和全基因组测序鉴定,分离获得1株西藏本土PPV,其基因组序列的大小为4336 bp,根据地域特点命名为PPV/China/XZ(XZ-PPV);进化树分析结果显示,XZ-PPV与国内参考株KX233726TJ、KJ201927和德国参考株JN4005167a、JN4005178a等PPV参考株处于同一分支,亲缘关系最近;生物信息学分析结果显示,XZ-PPV的NS1和VP2蛋白都属于亲水性稳定蛋白,二者均无信号肽序列和跨膜结构域,在二级结构中均以无规则卷曲为主要空间结构,且这2种蛋白的空间构象均相对稳定。综上所述,本试验发现XZ-PPV与目前已知PPV毒株的氨基酸亲水性位点和蛋白空间结构存在差异,如NS1和VP2蛋白存在亲水区域的氨基酸位点以及二级结构(α螺旋和β转角等)所占比例存有异同,可为后续PPV的诊断和相关疫苗研发提供数据支撑,也可为其防控和治疗提供一定的理论依据。
To elucidate the bioinformatic characteristics of the nonstructural protein 1(NS1)and viral structural protein 2(VP2)of Tibetan porcine parvovirus(PPV),this study conducted polymerase chain reaction(PCR)and full-genome sequencing on blood samples from suspected diseased Tibetan native pigs.Positive samples underwent virus isolation and cytopathic effect(CPE)observation.A phylogenetic tree of the isolated PPV strain was constructed to analyze genetic relatedness.Online tools and software were used to perform bioinformatic analysis of the amino acid sequences of NS1 and VP2,including their physicochemical properties and spatial structures.The results showed that one local PPV strain was successfully identified through PCR and whole-genome sequencing,with a genome length of 4336 bp.Based on regional origin,it was named PPV/China/XZ(XZ-PPV).Phylogenetic analysis indicated that XZ-PPV is most closely related to Chinese reference strains KX233726 TJ and KJ201927,as well as German strains JN4005167a and JN4005178a.Bioinformatic analysis revealed that both NS1 and VP2 of XZ-PPV are hydrophilic,stable proteins with no signal peptides or transmembrane domains.Their secondary structures are primarily composed of random coils,and the overall spatial conformations are relatively stable.In conclusion,XZ-PPV shows differences in amino acid hydrophilicity and protein spatial structure compared to previously known PPV strains.For example,differences and similarities exist in the amino acid sites within the hydrophilic regions of the NS1 and VP2 proteins,as well as in the proportions of their secondary structures(such asα-helices andβ-turns).These findings provide valuable data for the future diagnosis of PPV,vaccine development,and offer theoretical insights for prevention and treatment strategies.
作者
叶霄霞
罗侣
聂浩洋
刘福洲
李海洋
廉帅
纪菁蕊
韦明邦
商鹏
强巴央宗
YE Xiaoxia;LUO Lv;NIE Haoyang;LIU Fuzhou;LI Haiyang;LIAN Shuai;JI Jingrui;WEI Mingbang;SHANG Peng;QIANG Bayangzong(School of Animal Science,Xizang Agriculture and Animal Husbandry College,Nyingchi 860000,China;Key Laboratory of Tibetan Pig Genetic Improvement and Breeding,Nyingchi 860000,China;Provincial-Ministerial Collaborative Innovation Center for R&D of Agricultural and Animal Husbandry Resources with Tibetan Characteristics,Nyingchi 860000,China)
出处
《中国兽医杂志》
北大核心
2025年第8期53-61,共9页
Chinese Journal of Veterinary Medicine
基金
国家重点研发计划(2022YFD1600903)
国家自然科学基金地区科学基金项目(32160773)
西藏自治区科技计划项目(XZ202202JD0002N)
西藏自治区重大科技专项项目(XZ202101ZD0005N)。
