Identification of a collagen-like protein gene from white spot syndrome virus
Identification of a collagen-like protein gene from white spot syndrome virus
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DOI：
10.1007/s00705-003-0221-7
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发表时间：
2004-02-01
影响因子：
2.7
通讯作者：
Yang, F
中科院分区：
文献类型：
--
作者：
Li, Q;Chen, Y;Yang, F
白色斑点综合征病毒（WSSV）最独特的特征是存在胶原样蛋白（称为WSSV-CLP）。在这份报告中，WSSV-CLP（CLPn）的N-末端片段作为融合蛋白与谷胱甘肽S-转移酶（GST）在大肠杆菌中表达和纯化。然后针对纯化的融合蛋白（GST-CLPn）产生特异性抗体。时间序列分析表明，WSSV胶原基因是一个早期病毒基因。用特异性抗体进行的免疫金定位显示，在透射电子显微镜下，金颗粒沿着WSSV病毒粒子的外膜。本实验表明，胶原蛋白基因编码WSSV的包膜蛋白。用免疫亲和层析法从粗纯化的WSSV病毒粒子中纯化WSSV-CLP。WSSV-CLP被N-糖基化，如用N-连接糖苷酶F处理后SDS-PAGE中增加的迁移所示。
The most unique feature of white spot syndrome virus (WSSV) is the presence of a collagen-like protein (termed as WSSV-CLP). In this report, the N-terminal fragment of WSSV-CLP (CLPn) was expressed as a fusion protein with glutathione S-transferase (GST) in Escherichia coli and purified. Specific antibody was then raised against the purified fusion protein (GST-CLPn). Temporal analysis showed that the WSSV collagen gene was an early viral gene. Immunogold localization using specific antibody revealed that the gold particles, under a transmission electron microscope, were presented along the outer envelope of WSSV virions. This experiment suggested that the collagen gene encoded an envelope protein of WSSV. Using immuno-affinity chromatography, the WSSV-CLP was purified from crudely purified WSSV virions. The WSSV-CLP was N-glycosylated, as indicated by the increased migration in SDS-PAGE after treatment with N-linked glycosidase F.