2株水禽源坦布苏病毒的分离鉴定及全基因序列分析

doi:10.16431/j.cnki.1671-7236.2025.01.028

中国畜牧兽医 ›› 2025, Vol. 52 ›› Issue (1): 310-321.doi: 10.16431/j.cnki.1671-7236.2025.01.028

• 预防兽医 • 上一篇

2株水禽源坦布苏病毒的分离鉴定及全基因序列分析

陈作鑫^1,2, 潘彦林^1,3, 黄允真¹, 李林林¹, 董嘉文¹, 向勇¹, 徐志宏¹, 孙敏华¹, 黄淑坚², 廖明³, 张俊勤¹

1. 广东省农业科学院动物卫生研究所, 农业农村部禽流感等家禽重大疾病防控重点实验室, 广东省畜禽疫病防治研究重点实验室, 广东省动物疫病野外科学观测研究站, 广州 510640;
2. 佛山大学生命科学与工程学院, 佛山 528225;
3. 仲恺农业工程学院动物科技学院, 广州 510225

收稿日期:2024-05-10 发布日期:2024-12-30
通讯作者: 张俊勤 E-mail:Junqinzhang@yeah.net
作者简介:陈作鑫,E-mail:2506082851@qq.com。
基金资助:
国家自然科学基金(32102691);广州市科技计划项目(2024A04J6754);广东省畜禽疫病防治研究重点实验室项目(2023B1212060040);广东省动物疫病野外科学观测研究站项目(2021B1212050021)

Isolation,Identification and Whole Genome Sequence Analysis of Two Strains of Tambusu Virus from Waterfowl

CHEN Zuoxin^1,2, PAN Yanlin^1,3, HUANG Yunzhen¹, LI Linlin¹, DONG Jiawen¹, XIANG Yong¹, XU Zhihong¹, SUN Minhua¹, HUANG Shujian², LIAO Ming³, ZHANG Junqin¹

1. Institute of Animal Health, Guangdong Academy of Agricultural Sciences, Key Laboratory for Prevention and Control of Avian Influenza and Other Major Poultry Diseases, Ministry of Agriculture and Rural Affairs, Key Laboratory of Livestock Disease Prevention of Guangdong Province, Guangdong Provincial Observation and Research Station for Animal Disease, Guangzhou 510640, China;
2. College of Life Science and Engineering, Foshan University, Foshan 528225, China;
3. College of Animal Science & Technology, Zhongkai University of Agriculture and Engineering, Guangzhou 510225, China

Received:2024-05-10 Published:2024-12-30

摘要/Abstract

摘要： 【目的】探究水禽源坦布苏病毒(Tembusu virus,TMUV)流行毒株遗传变异情况及其膜前体蛋白(prM)和囊膜蛋白(E)的分子特征。【方法】利用仓鼠肾细胞(BHK-21)对疑似TMUV感染的鸭、鹅肝脏组织样品进行病毒分离,通过全基因克隆、测序、拼接获得分离株全基因序列。运用ModelFinder、MrBayes软件进行开放阅读框(open reading frame,ORF)、E基因的遗传进化分析;以GenBank数据库中TMUV MM1775株为参考,对分离株prM和E蛋白进行氨基酸突变位点分析;利用生物信息学在线软件对prM、E蛋白的跨膜区域、B细胞抗原表位、二级结构与三级结构进行预测。【结果】试验分离获得2株水禽源TMUV分离株,分别命名为FSE、ZJ2。全基因组遗传进化分析显示,分离株FSE、ZJ2分别属于Cluster 2.1.1与Cluster 2.2。氨基酸变异位点分析结果显示,FSE、ZJ2株prM和E蛋白中均存在多个氨基酸变异位点。生物信息学分析结果显示,FSE、ZJ2株E蛋白均存在2个跨膜区;prM、E蛋白中均存在多个线性B细胞抗原表位,与疫苗株DF-2的prM、E蛋白抗原表位氨基酸序列相比,FSE株prM蛋白存在E73H氨基酸位点突变,E蛋白存在A157V氨基酸位点突变;二级结构均以无规则卷曲为主,三级结构预测结果与二级结构相一致。【结论】本研究从广东地区分离得到2株Cluster 2 TMUV,2株分离株prM、E蛋白均存在多个氨基酸突变位点,表明TMUV在广东地区的流行过程中仍在不断发生变异。

关键词: 坦布苏病毒; 病毒分离; 遗传进化; 生物信息学

Abstract: 【Objective】 The objective of this experiment was to investigate the genetic variation of epidemic strains of Tembusu virus (TMUV) from waterfowl and the molecular characteristics of membrane precursor protein (prM) and membrane protein (E). 【Method】 BHK-21 cells were used to isolate the virus from the liver tissue samples of ducks and geese suspected to be infected with TMUV,and the whole gene sequence of the isolates was obtained by whole gene cloning,sequencing and splination.ModelFinder and MrBayes were used to analyze the genetic evolution of open reading frame (ORF) and E genes.With TMUV MM1775 in GenBank database as reference,amino acid mutation sites of prM and E proteins were analyzed.The transmembrane region,B-cell epitope,secondary and tertiary structures of prM and E proteins were predicted by bioinformatics online software. 【Result】 Two strains of TMUV from waterfowl were isolated and named as FSE and ZJ2,respectively.Whole genome genetic evolution analysis showed that the isolates FSE and ZJ2 belonged to Cluster 2.1.1 and Cluster 2.2,respectively.Amino acid variation sites analysis showed that there were multiple amino acid variation sites in prM and E proteins of the isolates FSE and ZJ2.The results of bioinformatics analysis showed that E protein of the isolates FSE and ZJ2 had 2 transmembrane regions.There were multiple linear B cell epitopes in prM and E proteins.Compared with the amino acid sequences of prM and E epitopes of vaccine strain DF-2,prM protein of the isolate FSE had mutation at the E73H amino acid site,and the E protein had mutation at the A157V amino acid site.The secondary structure was dominated by random coil,and the prediction result of tertiary structure was consistent with those of secondary structure. 【Conclusion】 In this study,two Cluster 2 TMUV were isolated from Guangdong province,and the prM and E proteins of the two isolates had multiple amino acid mutation sites,indicating that TMUV was still mutating during the epidemic process in Guangdong province.

Key words: Tembusu virus; virus isolation; genetic evolution; bioinformatics

中图分类号:

S852.65

陈作鑫, 潘彦林, 黄允真, 李林林, 董嘉文, 向勇, 徐志宏, 孙敏华, 黄淑坚, 廖明, 张俊勤. 2株水禽源坦布苏病毒的分离鉴定及全基因序列分析[J]. 中国畜牧兽医, 2025, 52(1): 310-321.

CHEN Zuoxin, PAN Yanlin, HUANG Yunzhen, LI Linlin, DONG Jiawen, XIANG Yong, XU Zhihong, SUN Minhua, HUANG Shujian, LIAO Ming, ZHANG Junqin. Isolation,Identification and Whole Genome Sequence Analysis of Two Strains of Tambusu Virus from Waterfowl[J]. China Animal Husbandry and Veterinary Medicine, 2025, 52(1): 310-321.

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2株水禽源坦布苏病毒的分离鉴定及全基因序列分析

Isolation,Identification and Whole Genome Sequence Analysis of Two Strains of Tambusu Virus from Waterfowl

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