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| Variation of Three Genes in White spot syndrome virus in Some Areas of China in 2017 |
| LIU Xiaoping1,2, CAI Miao1,2, LIU Qinghui1, WAN Xiaoyuan1, 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 and Rural Affairs, Qingdao Key Laboratory of Mariculture Epidemiology and Biosecurity, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China;
2. Shanghai Ocean University, Shanghai 201306, China |
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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.
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Received: 11 December 2018
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