【摘 要】
：
杆状病毒包埋型病毒粒子(occlusion-derived viruses,ODV)对昆虫宿主中肠上皮细胞有效识别和成功感染至少依赖8 种病毒编码的口服感染因子(per os infectivity factor,PIF)的协同作用,目前发现PIFs 在ODV 表面形成一个大的复合物(PIF complex),其中P74、PIF1-PIF4 是PIF complex 的组成成分,解析PIFcomp
【机 构】
：
中国科学院武汉病毒研究所病毒学国家重点实验室,湖北武汉430071
【出 处】
：
第六届全国农业微生物研究及产业化研讨会、第十五届全国杀虫微生物学术研讨会、第十一届全国虫生真菌学术研讨会
论文部分内容阅读
杆状病毒包埋型病毒粒子(occlusion-derived viruses,ODV)对昆虫宿主中肠上皮细胞有效识别和成功感染至少依赖8 种病毒编码的口服感染因子(per os infectivity factor,PIF)的协同作用,目前发现PIFs 在ODV 表面形成一个大的复合物(PIF complex),其中P74、PIF1-PIF4 是PIF complex 的组成成分,解析PIFcomplex 的组成和功能对于揭示杆状病毒的口服感染分子机理非常重要.本研究通过优化ODV 囊膜蛋白的提取方法,利用Blue-native PAGE 成功分离得到了苜蓿银纹夜蛾核多角体病毒(Autographa californica multiple nuclear polyhedrosis virus,AcMNPV)的PIF complex.结果显示：PIF complex 分子量大约为500 kDa；用前期制备的各种PIF 蛋白的抗体进行Western blot 分析,发现P74、PIF1、PIF2、PIF3、PIF6 和VP91 是PIF complex 的组分,而PIF5 和ODV-E66 不在PIF complex中；此外,构成PIF complex 的上述几种PIF 蛋白全部存在于复合物中,几乎检测不到游离的PIFs 存在.随后,利用一系列PIFs 单独缺失的病毒来研究各个PIF对复合物形成和功能的影响.Blue-native PAGE 和Western blot 分析显示,任何一个 PIF 的缺失均不同程度影响PIF complex 的形成.其中,PIF1 或PIF2 的缺失造成complex 完全消失；在PIF3 缺失的ODV 中,能检测一个包含PIF1 的分子量～200 kDa 的sub-complex 条带,当P74 或PIF6 缺失后,能检测到一个微弱的、分子量～300 kDa 的包含PIF1 和PIF2 的sub-complex；PIF4 缺失后,只能检测到包含VP91、PIF1 和PIF2 的分子量～400 kDa 的sub-complex.目前正在开展生物测定以及sub-complex 的组成解析,为深入揭示PIF complex 的形成机制和PIFs 的功能奠定基础.此外,生物信息分析显示在其他节肢动物病毒中,如salivary gland hypertrophy virus、white spot syndrome virus 能找到PIFs 的一些同源物(P74、PIF1-3),而nudivirus 还能进一步找到PIF5、PIF6 和VP91 的同源物,预示着口服感染是一个古老和不断演化的入侵机制.因此,揭示其他病毒中是否也存在相似的PIF complex 对于深入阐释这样一种古老病毒入侵机制也具有重要意义.
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