Cloning of Litopenaeus vannamei CD63 and it's role in white spot syndrome virus infection

被引:7
|
作者
Guan, Guang-Kuo [1 ,2 ]
Liu, Qing-Hui [1 ,3 ]
Li, Chen [1 ]
Huang, Jie [1 ,3 ]
机构
[1] Chinese Acad Fishery Sci, Yellow Sea Fisheries Res Inst, Qingdao, Peoples R China
[2] Shanghai Ocean Univ, Shanghai, Peoples R China
[3] Qingdao Natl Lab Marine Sci & Technol, Lab Marine Fisheries & Aquaculture, Qingdao, Peoples R China
来源
DEVELOPMENTAL AND COMPARATIVE IMMUNOLOGY | 2016年 / 60卷
基金
中国国家自然科学基金;
关键词
Litopenaeus vannamei; CD63; Cloning; WSSV; Interaction; TETRASPANIN FUNCTIONS; ENVELOPE PROTEINS; MAJOR ENVELOPE; SHRIMP; WSSV; VP28; IDENTIFICATION; PATHOGENESIS; DOMAINS; PCR;
D O I
10.1016/j.dci.2016.03.004
中图分类号
S9 [水产、渔业];
学科分类号
0908 ;
摘要
White Spot Syndrome Virus (WSSV) is currently the most serious shrimp pathogen, which has brought huge losses to shrimp industry worldwide. CD63 of shrimp belongs to the tetraspanin superfamily, which plays an important role in signal transduction and immune process. In this paper, CD63 cDNA sequence of Litopenaeus vannamei was cloned using RACE method. The amplified sequence is 1472 bp, with its ORF 744 bp, encoding 247 amino acids. Bioinformatics analysis showed that the sequence of LvCD63 has 93% similarity with Penaeus monodon and 92% similarity with Fenneropenaeus chinensis. Real-time PCR analysis showed that the mRNA levels of LvCD63 expressed in the tissues of hemocytes, gill, epithelial tissue, heart, lymphoid, hepatopancreas, stomach, intestines, muscle and nerve. Among these tissues the highest expression level was showed in the tissue of haemolymph, followed by epithelial tissue, hepatopancreas, and nerve. The lowest expression level of LvCD63 was appeared in the muscle tissue. After WSSV challenge, the expression levels of LvCD63 were both up-regulated in the tissues of gill and epithelial. However the expression level of LvCD63 in hepatopancreas was down-regulated. Far-western blot analysis showed that LvCD63 interacts with VP28, and both VP28N and VP28C fragments interact with LvCD63. Flow cytometry analysis showed that LvCD63 was present on the surface of hemocytes and it is required for binding of WSSV virions. Neutral experiments in vivo showed that LvCD63LEL delayed WSSV infection in shrimp. (C) 2016 Elsevier Ltd. All rights reserved.
引用
收藏
页码:209 / 217
页数:9
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