对虾白斑综合征病毒VP12的原核表达及其与VP24和VP26的相互作用  

Expression of White Spot Syndrome Virus VP12 and It's Interaction with VP24 and VP26

作　　者：马方方[1,2] 刘庆慧 黄倢[2] 

机构地区：[1]上海海洋大学,上海201306 [2]农业部海洋渔业资源可持续利用重点开放实验室中国水产科学研究院黄海水产研究所,山东青岛266071

出　　处：《动物医学进展》2013年第1期11-15,共5页Progress In Veterinary Medicine

基　　金：国家自然科学基金项目(30871942);国家虾产业体系项目(nycytx-46)

摘　　要：根据GenBank上对虾白斑综合征病毒(WSSV)囊膜蛋白基因VP12(wsv 432)的序列,设计并合成引物,PCR扩增得到VP12基因,构建重组表达载体pBAD/gⅢ–VP12并转化大肠埃希菌Top 10感受态细胞。基因工程菌株在37℃用L-阿拉伯糖诱导,表达产物经Western blot和SDS-PAGE检测显示有与预期大小19ku相符合的目的蛋白。以Co2+亲和层析方法,获得纯化VP12蛋白。采用Far-Western blot结合ELISA方法,分析VP12与WSSV结构蛋白的作用。结果表明,VP12与囊膜蛋白VP24、VP26具有相互作用,该研究对于揭示WSSV分子组装机制,阐明其致病机理具有重要意义。A pair of primers were designed according to the sequence of VP12 gene of White spot syndrome virus （WSSV） in GenBank. The VP12 DNA fragment was amplified by PCR and cloned into E. coli ex- pression vector pBAD/gIII successfully. Then pBAD/gIII-VP12 was transformed into E. coli- Top 10 cells. After L-arabinose induction at 37 ℃, the fusion protein with 19 ku was expressed, which was con- formed by SDS-PAGE and Western-blot analysis. The envelope protein VP12 was purified by Co2＋-column affinity chromatography. The interaction between purified protein VP12 and WSSV structure protein was analyzed by Far-western blot and ELISA. It is concluded that VP12 could interact with VP24 and VP26, which had important significance to reveal WSSV molecule assembly system and clarify WSSV pathogene- sis mechanism.

关 键 词：对虾白斑综合征病毒 VP12 原核表达 VP24 VP26 相互作用 

分 类 号：S945.19[农业科学—水产养殖] Q786[农业科学—水产科学]