对虾白斑综合征病毒非结构蛋白ICP11的研究

The Research of White Spot Syndrome Virus Nonstructural Protein ICP11

白斑综合征病毒(WSSV)是一种对虾养殖业中危险极大的传染性病原体,它是一种双链环状DNA杆状型病毒.ICP11是WSSV感染宿主后产生的一种高丰度表达蛋白,根据icp11基因的序列,通过PCR扩增获得了该全长基因,将其克隆在载体pET-His上,以E.coliBL21为宿主菌,成功表达并纯化了含His标记的目的蛋白,并制备了特异性鼠抗血清.Western blot杂交实验结果显示,该蛋白不存在于纯化的病毒粒子的结构蛋白中,只存在于病毒感染的虾中肠组织的总蛋白中.这说明ICP11是WSSV的一种非结构蛋白.通过Far-western blot杂交分析,发现重组表达的ICP11会发生自身蛋白的聚合作用.凝胶过滤色谱层析法进一步证实ICP11蛋白可以聚合为二聚体.

White spot syndrome virus （WSSV）, a rod-shaped double-stranded circular DNA virus,is a kind of major dangerous .infectious pathogen in shrimp culture industry. ICPll（Infected cell protein 11）was generated from a high abundance protein in WSSV-infected host. According to icp11 gene sequence, the full-length gene was amplified by PCR and then cloned in vector pET-His. The gene with 6 His-tag was expressed and purified, then was used to prepare specific antibody. Western blot results revealed that the protein is not present in the strueture proteins of purified virions, but only in WSSV-infected shrimp total protein extracted from mid- gut tissue. It is suggested that ICP11 is a WSSV nonstruetural protein. ICP1l was found being capable of self-interaction through Farwestern blot analysis, and capable of self-multimerization for a dimer using the method of gel filtration chromatography.