【摘要】 本实验利用 AcMNPV(Autographa californica multiple nuclear polyhedrosis virus,AcMNPV)的 bac-to-bac 系统构建了两种重组病毒,即含 GFP-actin 融合基因的重组病毒 AcMNPV-GFP-actin 和含 GFP 基因的重组病毒AcMNPV-GFP。用这两种重组病毒分别感染 Sf9 细胞,以 AcMNPV-GFP 感染的 Sf9 细胞为对照,用共聚焦激光扫描显微镜观察了绿色荧光在病毒感染过程中的分布情况。由于肌动蛋白和绿色荧光蛋白是共定位的,所以绿色荧光的分布情况就是肌动蛋白的分布情况。实验中观察发现,AcMNPV-GFP 感染的 Sf9 细胞中的绿色荧光,在整个感染过程中是弥散分布的,而 AcMNPV-GFP-actin 感染 Sf9 细胞后 24-72h 这段时间内,肌动蛋白最初聚集在细胞核内,随后逐渐由细胞核向细胞质转移,最后完全聚集于细胞膜。根据实验结果,推测肌动蛋白可能参与了AcMNPV 出芽型病毒粒子(BV)由细胞核向细胞质运输以及从细胞膜排出的过程。更多还原

【Abstract】 To investigate the role of actin in AcMNPV BV transportation from nuclear to cytoplasm, we constructed two recombinant viruses AcMNPV-GFP and AcMNPV-GFP-actin, in which the expression of GFP gene and GFP-actin fusion gene was driven by polyhedrin promoter. Here, we report the expression pattern of GFP in Sf9 cells infected with AcMNPV-GFP and AcMNPV-GFP-actin. Within 24-72 hr after infection, we observed that GFP signal in Sf9 cells infected with AcMNPV-GFP-actin was first concentrated in nuclear and then gradually located to cytoplasm and finally accumulated exclusively at cytoplasmic membrane. In the Sf9 cells infected with AcMNPV-GFP, however, the GFP signal was diffused throughout cell and this expression pattern did not change during the same period of observation. Thus, our results indicated that actin might participate in the transportation of the AcMNPV BV from nuclear to cytoplasm and its exocytosis from the cell.更多还原