Zhu Yanbing, Xie Xixian, Yang Feng
School of Life Science, Xiamen University, Xiamen 361005, People's Republic of China.
Virus Res. 2005 Nov;113(2):100-6. doi: 10.1016/j.virusres.2005.04.020.
White spot syndrome virus (WSSV) is one of the most virulent pathogens in shrimp culture worldwide. Combining SDS-PAGE with mass spectrometry, a novel envelope protein from WSSV was identified to match an open reading frame (ORF) of WSSV genome. This ORF contained 3582nt, encoding 1194 aa, and was termed the vp124 gene. One part of the whole gene (named vp124p) was cloned into pET-GST vector and expressed as a fusion protein with glutathione S-transferase (GST) in Escherichia coli strain BL21 (DE3). Specific antibodies were raised using the purified fusion protein (GST-VP124P). Temporal transcription analysis revealed that the vp124 gene was a late gene. Western blot analysis showed that the mouse anti-GST-VP124P antibodies reacted specifically with VP124 present either in the WSSV virions or in the viral envelopes, and did not react with the proteins of the viral nucleocapsids. VP124 was located in the WSSV virions as an envelope protein using immunoelectron microscopy.
白斑综合征病毒（WSSV）是全球对虾养殖中最具毒性的病原体之一。通过将十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）与质谱分析相结合，鉴定出一种来自WSSV的新型包膜蛋白，它与WSSV基因组的一个开放阅读框（ORF）相匹配。该ORF包含3582个核苷酸，编码1194个氨基酸，被命名为vp124基因。将整个基因的一部分（命名为vp124p）克隆到pET-GST载体中，并在大肠杆菌BL21（DE3）菌株中作为与谷胱甘肽S-转移酶（GST）的融合蛋白进行表达。使用纯化的融合蛋白（GST-VP124P）制备特异性抗体。时间转录分析表明vp124基因是一个晚期基因。蛋白质印迹分析显示，小鼠抗GST-VP124P抗体与WSSV病毒粒子或病毒包膜中存在的VP124特异性反应，而不与病毒核衣壳蛋白反应。使用免疫电子显微镜确定VP124作为包膜蛋白位于WSSV病毒粒子中。
