Chen Li-Li, Lu Li-Chen, Wu Wan-Jung, Lo Chu-Fang, Huang Wei-Pang
Department of Biomedical Sciences, Chung Shan Medical University, No. 110, Sec. 1, Chien-Kuo N. Road, Taichung 402, Taiwan, ROC.
Dis Aquat Organ. 2007 Mar 13;74(3):171-8. doi: 10.3354/dao074171.
White spot syndrome virus (WSSV) is the causative agent of a severe disease of cultivated shrimp. Using purified WSSV virions, VP53A encoded by open reading frame wssv067 was identified as a structural protein by SDS-PAGE and proteomics. Immunoelectron microscopy with a gold-labeled secondary antibody revealed that VP53A was distributed on the viral envelope. In order to further explore the link between WSSV067 and host proteins, we performed a yeast 2-hybrid screening of a Penaeus monodon cDNA library, using WSSV067C as bait. One of the molecules that specifically interacted with WSSV067C was the P. monodon chitin-binding protein (PmCBP). An in vitro binding assay showed that c-myc-WSSV067C was capable of co-precipitating HA-PmCBP-C. Furthermore, PmCBP was expressed in almost all organs but appeared to be up-regulated at the late stage of WSSV infection.
白斑综合征病毒（WSSV）是养殖虾严重疾病的病原体。使用纯化的WSSV病毒粒子，通过SDS-PAGE和蛋白质组学鉴定出开放阅读框wssv067编码的VP53A为结构蛋白。用金标记的二抗进行免疫电子显微镜检查显示，VP53A分布在病毒包膜上。为了进一步探索WSSV067与宿主蛋白之间的联系，我们以WSSV067C为诱饵，对斑节对虾cDNA文库进行了酵母双杂交筛选。与WSSV067C特异性相互作用的分子之一是斑节对虾几丁质结合蛋白（PmCBP）。体外结合试验表明，c-myc-WSSV067C能够共沉淀HA-PmCBP-C。此外，PmCBP在几乎所有器官中都有表达，但在WSSV感染后期似乎上调。
