China Animal Husbandry and Veterinary Medicine ›› 2025, Vol. 52 ›› Issue (1): 310-321.doi: 10.16431/j.cnki.1671-7236.2025.01.028

• Preventive Veterinary Medicine • Previous Articles    

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

CHEN Zuoxin1,2, PAN Yanlin1,3, HUANG Yunzhen1, LI Linlin1, DONG Jiawen1, XIANG Yong1, XU Zhihong1, SUN Minhua1, HUANG Shujian2, LIAO Ming3, ZHANG Junqin1   

  1. 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

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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

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