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2015年中国典型对虾养殖区WSSV流行株高变异区序列的分析比较
秦梦雪1,2, 孙新颖1, 万晓媛1, 刘庆慧1, 黄 倢1
1.青岛海洋科学与技术国家实验室海洋渔业科学与食物产出过程功能实验室 农业部海水养殖病害防治重点实验室 青岛市海水养殖流行病学与生物安保重点实验室 中国水产科学研究院黄海水产研究所 青岛 266071;2.上海海洋大学水产与生命学院 上海 201306
摘要:
为了解我国白斑综合征病毒(WSSV)流行变异特征,本研究对2015年4~10月期间在山东、江苏、天津、浙江、海南和广东6省市采集到的57份WSSV阳性的样本,通过特异性的扩增目的片段,根据测序结果分析比较不同地区、不同分离株之间在ORF14/15、ORF23/24上的缺失变异情况,以及ORF75、ORF94和ORF125上的重复单元(Repeat unit, RU)数目差异。结果显示,在ORF14/15扩增中,分别有6530、5908和5725 bp的片段缺失,而在ORF23/24扩增中均有12070 bp大片段的缺失,ORF75的45 bp的RU数目分别为1、2和3,102 bp的RU数目均为1,而ORF94的RU数目分别为4、5、10和12不等,ORF125的RU数目为3、5和6。结果表明,WSSV在部分开放型阅读框上表现出明显的变异差异,而在某些开放型阅读框上的缺失情况则有显著的稳定性。
关键词:  WSSV  ORF  缺失  变异
DOI:
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基金项目:
Variable Sequence Comparison of WSSV Regions from Different Parts of China in 2015
QIN Mengxue1,2, SUN Xinying1, WAN Xiaoyuan1, LIU Qinghui1, HUANG Jie1
1.Laboratory for Marine Fisheries Science and Food Production Processes, Qingdao National Laboratory for Marine Science and Technology, Key Laboratory of Maricultural Organism Disease Control, Ministry of Agriculture, Qingdao Key Laboratory of Mariculture Epidemiology and Biosecurity, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Science, Qingdao 266071;2.College of Fisheries and Life Science, Shanghai Ocean University, Shanghai 201306
Abstract:
In recent years, the fast development of aquaculture has been followed with the increased diseases such as the white spot disease to cause a great loss for the global aquaculture industry. To understand the variations in WSSV ORF14/15 and ORF23/24 and the VNTR in different regions of China in 2015, we investigated 57 samples WSSV-positive on PCR, collected in a disease outbreak area between April and October 2015. The tested samples were from Shandong, Guangdong, Hainan, Zhejiang, Tianjin and Jiangsu. Using specific primers, WSSV-positive samples were genotyped and the amplified fragments were ligated to a T-vector and transformed in Top10 cells. The positive clones were selected and sequenced. After sequencing, the fragments deleted from ORF14/15 and ORF23/24 in different samples were compared with the sequences of Th-96-Ⅱ and TW respectively, and three WSSV VNTR regions in ORF75, ORF94, and ORF125 from different samples were analyzed with the DNAMAN software. The results showed that 35 samples had the products of ORF14/15 amplification, and 14 sample had the products of ORF23/24 amplification. The lengths of the amplified fragments of ORF14/15 were 1270 bp, 1892 bp and 2075 bp, corresponding to deletions of 6530 bp, 5908 bp and 5725 bp, respectively, compared with Th-96-Ⅱ. One type of ORF23/24 was confirmed, and the amplified fragment was 1140 bp, corresponding to a deletion of 12070 bp compared with the TW isolate. Ten fragments were amplified from ORF75, including 1, 2, 3 repeat units with 45 bp, and 1 with 102 bp. Four different ORF94 VNTRs, including 4, 5, 10, or 12 repeat units, were identified. The ORF125 VNTR including 3, 5 or 6 repeat units was identified. The results showed that WSSV exhibited a significant difference in some open reading frames, but a significant stability in others.
Key words:  WSSV  ORF  Deletion  Variation