2017年中国部分地区白斑综合征病毒3个基因的变异分析

doi:10.16378/j.cnki.1003-1111.2020.04.013

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摘要白斑综合征病毒广泛分布于中国、东南亚、南亚、南美洲、北美洲的许多国家,并且具有较高的致病性。为探明中国不同地区白斑综合征病毒毒株的变异情况,本研究选取2017年2—6月在河北、浙江、山东、上海、福建和广东共6个省市的白斑综合征病毒病害暴发区的44份患病样本,提取其DNA,进行wsv-pol、wsv313、wsv150的PCR扩增,筛选阳性克隆并进行测序分析。分析结果显示,在2017年wsv-pol的12个阳性样本中,均在第164位出现单个甘氨酸缺失,在个别样品中也存在氨基酸的替换现象;在wsv313的PCR扩增中有9个阳性样本在第280位插入1个脯氨酸,2个样本在第64位天冬氨酸突变为甘氨酸;wsv150的阳性样本均在第587位插入碱基T。氨基酸比对结果显示,8个样本在第200位之后出现106个大片段氨基酸的缺失,同时在第196、197、198、199位发生氨基酸的替换。氨基酸比对结果表明,wsv-pol、wsv313均未出现较大程度的变异,而wsv150的氨基酸序列出现了较大程度的变异,但未表现出地区差异性。本研究将为白斑综合征病毒病原学的研究提供依据。

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关键词 ：白斑综合征病毒, wsv-pol, wsv313, wsv150, 序列分析

Abstract：White spot syndrome virus (WSSV), as a high pathogenic agent, is widely prevalent in China, Southeast Asian, India and South America. In this study, 44 WSSV-positive individuals were collected from the area of Hebei province, Zhejiang province, Shandong province, Shanghai, Fujian province and Guangdong province from February to June of 2017 and DNA of the samples was extracted and specific primers for three pairs of variable regions were designed to understand the variation of the WSSV. The samples were subjected to PCR amplification of wsv-pol, wsv313, and wsv150, and the specifically amplified bands were purified using a gel recovery kit, ligated into the pMD^©18-T vector, transformed into DH5α competent cells, screened for positive clones and sequenced. The results showed that there were a total of 12 positive samples in the amplification of wsv-pol, with detection rate of 27%. The positive samples all showed a single glycine deletion at position 164, and there were also amino acid substitutions in individual samples. Amino acid alignments showed that there was no significant variation in wsv-pol. For wsv313, twelve positive samples were also found in the PCR amplification. Among the 9 positive samples, a proline was inserted at the 280th position. There were two samples which the aspartic acid was mutated to glycine at the position of 64.There were a total of 8 positive samples in the amplification of wsv150, with detection rate of some 18%. Analysis of its nucleotide sequence revealed that the positive samples were only inserted base T at position of 587. The amino acid sequence alignment showed that the positive samples all had 106 large fragment amino acid deletions posterior to the 200th position, with amino acid substitutions at positions 196, 197, 198, and 199. The amino acid sequence showed a large degree of variation, without regional differences. The findings provided the data for etiology of WSSV.

Key words： WSSV wsv-pol wsv313 wsv150 sequence analysis

收稿日期: 2018-12-11

PACS:

Q78

基金资助:国家自然科学基金资助项目(31672679);中国水产科学研究院黄海水产研究所基本科研业务费项目(20603022017017).

通讯作者: 刘庆慧(1962—),女,研究员;研究方向:对虾病原生物学.E-mail:liuqh@ysfri.ac.cn.

作者简介: 刘晓娉(1994—),女,硕士研究生;研究方向:水生生物学. E-mail:1197098046@qq.com.

引用本文:

刘晓娉, 蔡苗, 刘庆慧, 万晓媛, 黄倢. 2017年中国部分地区白斑综合征病毒3个基因的变异分析[J]. 水产科学, 2020, 39(4): 560-566.
LIU Xiaoping, CAI Miao, LIU Qinghui, WAN Xiaoyuan, HUANG Jie. Variation of Three Genes in White spot syndrome virus in Some Areas of China in 2017. Fisheries Science, 2020, 39(4): 560-566.

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http://www.shchkx.com/CN/10.16378/j.cnki.1003-1111.2020.04.013 或 http://www.shchkx.com/CN/Y2020/V39/I4/560

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