Chen Guoqing, Fang Yang, Wu Lijuan, Yan Qing, Krell Peter J, Feng Guozhong
State Key Laboratory of Rice Biology, China National Rice Research Institute, Hangzhou, 311400, China.
China National Rice Research Institute, Hangzhou, 311400, China.
Arch Virol. 2017 Nov;162(11):3487-3492. doi: 10.1007/s00705-017-3468-0. Epub 2017 Jul 20.
DNA polymerase (DNApol) is present in all baculoviruses and plays a crucial role in viral DNA replication. Previously we showed that the DNApol of the alphabaculovirus group II Spodoptera litura nucleopolyhedrovirus (SpltNPV) could partially substitute for the DNApol of a group I alphabaculovirus, Autographa californica multiple nucleopolyhedrovirus (AcMNPV). However, it is not known if a betabaculovirus DNApol could subsititute for the alphabaculovirus DNApol in AcMNPV. In this report, DNApol of the betabaculovirus Pieris rapae granulovirus (PiraGV) was inserted into a dnapol-null AcMNPV bacmid, creating Bac-AcΔpol:PrPol. The repair virus did not spread to neighboring cells; virus growth curve and real-time PCR revealed that the PiraGV dnapol substitution abrogated AcMNPV DNA replication and virus production. Immunofluorescence microscopy showed that PiraGV DNApol could be expressed and localized to the nucleus. Collectively, our results suggested that the alphabaculovirus AcMNPV DNApol could not be replaced by a DNApol from the betabaculovirus, PiraGV.
DNA聚合酶(DNApol)存在于所有杆状病毒中,并在病毒DNA复制中起关键作用。此前我们表明,II组甲型杆状病毒斜纹夜蛾核多角体病毒(SpltNPV)的DNApol可以部分替代I组甲型杆状病毒苜蓿银纹夜蛾多粒包埋核型多角体病毒(AcMNPV)的DNApol。然而,尚不清楚乙型杆状病毒的DNApol是否可以替代AcMNPV中的甲型杆状病毒DNApol。在本报告中,将乙型杆状病毒菜青虫颗粒体病毒(PiraGV)的DNApol插入到缺失dnapol的AcMNPV杆粒中,构建出Bac-AcΔpol:PrPol。修复病毒未扩散到邻近细胞;病毒生长曲线和实时PCR显示,PiraGV的dnapol替代作用消除了AcMNPV的DNA复制和病毒产生。免疫荧光显微镜检查表明,PiraGV DNApol可以表达并定位于细胞核。总体而言,我们的结果表明,甲型杆状病毒AcMNPV的DNApol不能被乙型杆状病毒PiraGV的DNApol所取代。