对虾白斑综合症病毒核衣壳蛋白VP664的鉴定研究

Study of white spot syndromes virus nucleocapsid protein VP664

对虾白斑综合症病毒核衣壳蛋白VP664是至今知道的分子量最大的结构蛋白,全长18 234bp,预计编码6 077个氨基酸.从编码vp664基因的两端各选600bp,分别命名为vp664n、vp664c进行原核表达,成功表达纯化了目的蛋白并制备抗体.蛋白印记杂交(Western blot)实验中蛋白特异抗体只与完整的病毒和核衣壳裂解蛋白中的VP664蛋白反应,而不和膜蛋白反应,证明VP664基因所编码的蛋白在完整病毒颗粒上定位于核衣壳.

White spot syndromes virus nucleocapsid protein VP664 is a hitherto biggest structural protein. The genome is the presence of a giant open reading frame（ORF） of 18 234 nucleotids that encodes a long polypeptide of 6 077 amino acids. Two 600bp were chosen from two sides of gene that encoded vp664 , and correspondingly each was given the name vp664n.vp664c.They were separate cloned and inserted into pET-his vector , The gene was expressed in E. coli BL21 （DE3） and purified, then was used to prepare specific antibody. The antibodies of this protein did only react with the proteins of WSSV nucleocapsids and the intact virions in western blot analysis except for wanberance protein , indicating that this protein should belong to viral nucleocapsids