绿色荧光示踪马传染性贫血病毒样颗粒在活细胞中的组装过程  

Tracing the assembly process of GFP-taged equine infectious anemia virus-like particles in live cells

作　　者：张泽力[1,2] 马建 尹鑫[2,3] 谷勤雍[2,3] 艾有为[1,2] 曹苏亚[1,2] 王玉龙[1] 王晓钧[2] 

机构地区：[1]东北林业大学野生动物资源学院,黑龙江哈尔滨150040 [2]中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室/大动物研究室,黑龙江哈尔滨150001 [3]东北农业大学动物医学院,黑龙江哈尔滨150030

出　　处：《中国预防兽医学报》2013年第9期691-694,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家自然基金项目(31072113);兽医生物技术国家重点实验室基金项目(SKLVBP201205;201304)

摘　　要：马传染性贫血病毒(EIAV)gag基因编码的多聚Gag蛋白能够自动组装形成病毒样颗粒(VLPs)。为研究EIAV衣壳蛋白合成、组装及释放过程,本实验通过构建表达EIAV Gag-GFP融合蛋白重组质粒,并将其转染293T细胞,采用western blot检测不同时间点VLPs的产生及释放。并进一步通过活细胞和3D成像技术观察Gag-GFP在细胞中的表达动态及VLPs的组装过程。结果表明:Gag和GFP蛋白融合表达不影响VLPs的组装,转染约6 h后EIAV Gag蛋白进行合成并在细胞胞质聚集完成组装形成VLPs。该实验结果为进一步研究EIAV在细胞中的组装路线和机制奠定了重要基础。Gag polyprotein of equine infectious anemia virus （EIAV） has the ability of auto-assembling to virus-like particles （VLPs）. To investigate the assembly process of EIAV in live cells, the eukaryotic expression recombinant plasmid of pcDNA-Gag- GFP was constructed, and the Gag-GFP was expressed in 293T cells transfected with the recombinant plasmid and the production of VLPs was detected in different time points by using western blot. Additionally, the dynamic expression of Gag-GFP and assembly process of VLPs were traced by live cell and 3D imaging technology. The results showed that the fusion expression of Gag with GFP protein was not impacted the formation of VLPs. Moreover, the Gag-GFP was expressed and then completed assembly of the VLPs in 293T cell after 6 hours transfection with pcDNA-Gag-GFP and it was also revealed that EIAV Gag protein multimerizined in the cell cytoplasm to assembly of VLPs. This experiment laid the foundation for further study of the assembly sites and mechanism of EIAV.

关 键 词：马传染贫血病毒 病毒样颗粒 活细胞共聚焦 病毒组装 

分 类 号：S852.65[农业科学—基础兽医学]