肌动蛋白抑制了杆状病毒多角体蛋白基因的转录与表达  

Actin and Production of AcMNPV Polyhedra

作　　者：贾蕴莉[1,2] 余泽华[2] 陈新文[1] 

机构地区：[1]中国科学院武汉病毒研究所,病毒学国家重点实验室,武汉430071 [2]华中师范大学生命科学院昆虫研究所,武汉430079

出　　处：《生物化学与生物物理进展》2005年第9期829-834,共6页Progress In Biochemistry and Biophysics

基　　金：国家重点基础研究发展规划项目(973)(2003CB1140);国家自然科学基金资助项目(30325002;39870039).~~

摘　　要：为进一步研究肌动蛋白在杆状病毒感染晚期的作用,利用Bac-to-Bac系统构建了表达多角体基因、肌动蛋白与绿色荧光蛋白融合基因的重组病毒vAc-ph/70GA,同时构建对照病毒vAc-ph.实验发现,重组病毒vAc-ph/70GA感染Sf9细胞后能持续表达肌动蛋白,但不能形成多角体,vAc-ph则能形成多角体.SDS-聚丙烯酰胺凝胶电泳(SDS-PAGE)显示,vAc-ph/70GA感染晚期,细胞内没有多角体蛋白的表达;RT-PCR的结果进一步表明多角体基因的转录被抑制.肌动蛋白的表达并没有影响vAc-ph/70GA对细胞的感染力.结果表明,晚期表达的外源肌动蛋白抑制了多角体基因的转录和表达,从而导致多角体不能正常产生.Studies have shown that host actin is essential for baculovirus replication and assemblage. In order to further elucidate the function of actin in the late and very late infection stages, a recombinant virus vAc-ph/70GA were constructed using Bac-to-Bac system, in which eGFP-actin fusion gene under control of hsp70 promoter and polyhedrin gene （ph） with its＇ own promoter were inserted, vAc-ph, which only contains ph, was also constructed as control. After vAc-ph/70GA infected Sf9 cells, actin was expressed persistently. However, there were no polyhedra, while the control virus vAc-ph did produce polyhedra. Further analysis such as SDS-PAGE and RT-PCR did not detect the transcription and expression of polyhedrin after vAc-ph/70GA infected Sf9 cells, indicating that the late expression of actin inhibit polyhedra formation. Expression of actin, however, did not change the viral infectivity. It can be concluded that the persistently expression of actin during late stage of baculovirus infection inhibit the polyhedrin expression, and then the formation of polyhedra. Here, some results were shown by electro microscopy, SDS-PAGE and RT-PCR with vAc-ph/70GA.

关 键 词：苜蓿银纹夜蛾多核衣壳核型多角体病毒 肌动蛋白 多角体 多角体基因 

分 类 号：Q7[生物学—分子生物学]