【摘要】 苜蓿尺蠖核型多角体病毒 (Autographacalifornicanuclearpolyhedrosisvirus ,AcMNPV)在细胞质中合成其囊膜蛋白 ,但在细胞核内组装并包埋病毒粒子 ,这些蛋白的核定向转运机制是人们甚感兴趣的课题。以AcMNPV多角体衍生型病毒ODV(occlusion -derivedvirus ,ODV)的一种囊膜蛋白ODV -E18为对象 ,通过E18与一个标记短肽Flag融合的重组病毒的构建 ,以免疫荧光法跟踪检测E18蛋白的转运过程及形态 ,并利用酵母双杂交系统法(yeasttwohybridsystem) ,通过蛋白 -蛋白相互作用的研究 ,寻找与E18紧密结合的可能的转运蛋白。研究结果表明 ,E18是先以核内模结构———微泡 (microvesicle)的形式存在于核内的 ;一个被报道具有核定向转运功能的AcM NPV囊膜蛋白 -ODV -E6 6能与E18形成紧密的复合体 ,推测E - 6 6可能在E18的核定向转运中起着运载体的作用。更多还原

【Abstract】 The envelope proteins of Autographa californica nuclear polyhedrosis virus (AcMNPV) occlusion-derived virus are synthesized in cytosolic plasma of their host cells, transported and assembled in the nuclei of the cells. The mechanism of protein transporting and targeting is of quite interest to us. In this study, a recombinant virus expressing the fusion protein of E18 and Flag marker peptide under AcMNPV polyhedrin promoter was constructed. Immunofluorescence was used to localize E18 in the recombinant virus infected cells and yeast two hybrid system was employed to find out the possible transporting protein which binds with E18. Our preliminary results showed that E18 existed first in the nuclei in the form of microvesicle-the intranuclear membrane structure and it might be transported into the nuclei through the interaction with another AcMNPV envelope protein-ODV-E66, the latter has been reported containing a 23-amino-acid sequence which is sufficient to direct reporter proteins to nucleus.更多还原