浙江农业学报 ›› 2022, Vol. 34 ›› Issue (11): 2368-2378.DOI: 10.3969/j.issn.1004-1524.2022.11.06

• 动物科学 • 上一篇    下一篇

一株牦牛源牛病毒性腹泻病毒毒株的分离鉴定及其遗传特性分析

王慧慧1,2(), 冯茜莉1,2, 汪梦竹1,2, 赵泽阳1,2, 李易聪1,2, 蒲飞洋1,2, 马鹏1,2, 李勇3, 龚真莉3, 马忠仁1, 马晓霞1,*()   

  1. 1.西北民族大学 生物医学研究中心,甘肃 兰州 730030
    2.西北民族大学 生命科学与工程学院,甘肃 兰州 730010
    3.中国农业科学院 兰州兽医研究所,甘肃 兰州 730046
  • 收稿日期:2021-10-28 出版日期:2022-11-25 发布日期:2022-11-29
  • 通讯作者: 马晓霞
  • 作者简介:*马晓霞,E-mail: maxiaoxia956@163.com
    王慧慧(1997—),女,甘肃天水人,硕士研究生,研究方向为病毒生物分子与免疫学。E-mail: 3407975334@qq.com
  • 基金资助:
    校企横向合作项目(2021西民合(科研)字第175号)

Isolation, identification and analyses for genetic features of one bovine viral diarrhea virus strain from yak

WANG Huihui1,2(), FENG Xili1,2, WANG Mengzhu1,2, ZHAO Zeyang1,2, LI Yicong1,2, PU Feiyang1,2, MA Peng1,2, LI Yong3, GONG Zhenli3, MA Zhongren1, MA Xiaoxia1,*()   

  1. 1. Biomedical Research Center, Northwest Minzu University, Lanzhou 730030, China
    2. College of Life Science and Engineering, Northwest Minzu University, Lanzhou 730010, China
    3. Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, China
  • Received:2021-10-28 Online:2022-11-25 Published:2022-11-29
  • Contact: MA Xiaoxia

摘要:

通过病毒分离培养技术,从牦牛血清中分离获得一株致细胞病变的牛病毒性腹泻病毒(BVDV),命名为GSTZ毒株。经电镜观察,GSTZ毒株的直径在50~60 nm。按Karber法测算,该病毒滴度为5.0×106.5 mL-1[以组织半数感染量(TCID50)计]。利用反转录PCR(RT-PCR)测定GSTZ毒株的完整开放阅读框(ORF),全长11 691 nt,将其与参考毒株ORF在核酸序列和同义密码子使用模式等方面进行分析,确定GSTZ毒株在遗传学特性上属于基因1型家族成员。在GSTZ毒株的完整ORF中,A+U的出现频率要高于G+C,而且,病毒同义密码子的使用模式体现出对U/A结尾同义密码子使用偏嗜性高的遗传学特征。GSTZ毒株明显降低了使用含有CpG二联核苷酸的同义密码子的频率。这有助于降低病毒对宿主细胞免疫系统的刺激强度,从而促进病毒的复制增殖。研究成果可为进一步研究BVDV的相关分子机制提供试验材料,并为BVDV不同基因型的抗原关系分析与BVDV疫苗的研制奠定基础。

关键词: 牛病毒性腹泻病毒, 牦牛, 基因1型, 同义密码子, 致细胞病变

Abstract:

In this study, one bovine viral diarrhea virus (BVDV) strain was successfully isolated and termed as GSTZ. By electron microscopy, several 50-60 nm viral particles were observed. GSTZ strain displayed the TCID50 (50% tissue culture infection dose) value of 5.0×106.5 mL-1 as revealed by Karber method. Via reverse transcription-PCR (RT-PCR), the whole open reading frame (ORF) of GSTZ strain was sequenced (whole length of 11 691 nt) and classified into the genotype 1 group at either nucleotide level or synonymous codon usage level. The appearance frequency of A+U was higher than that of G+C in GSTZ ORF, and the synonymous codon usage pattern of GSTZ strain displayed the obvious usage bias in synonymous codons with U/A. It was worth noting that, this strain had a tendency of avoiding the selection of the synonymous codons containing CpG dinucleotides, which strongly suggested that the GSTZ strain might weaken the immune responses from host in order to successfully generate its viral particles. The isolated BVDV strain and the above findings could provide biological materials for the related mechanism study of BVDV, and lay foundations for the analysis of antigen relationship of different genotypes of BVDV and vaccine development.

Key words: bovine viral diarrhoea virus, yak, genotype 1, synonymous codon, cytopathogenic effect

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