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

作　　者：熊生良[1] 杨丰[2] 

机构地区：[1]厦门大学生命科学学院,福建厦门361005 [2]国家海洋局第三海洋研究所,福建厦门361005

出　　处：《台湾海峡》2007年第1期46-52,共7页Journal of Oceanography In Taiwan Strait

基　　金：国家自然科学基金资助项目(30330470);福建省自然科学基金资助项目(2003F001)

摘　　要：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

关 键 词：分子生物学 蛋白印记杂交(Western blot) 表达纯化 核衣壳蛋白 对虾白斑 综合症病毒 

分 类 号：Q945.19[生物学—植物学]