Berretta Marcelo F, López M Gabriela, Taboga Oscar, Sciocco-Cap Alicia, Romanowski Víctor
Instituto de Microbiología y Zoología Agrícola, INTA Castelar, Castelar, Argentina.
Virus Genes. 2013 Feb;46(1):152-61. doi: 10.1007/s11262-012-0843-5. Epub 2012 Nov 4.
We used transient expression assays to assess the function of the baculovirus Spodoptera frugiperda M nucleopolyhedrovirus (SfMNPV) homologs of Autographa californica MNPV (AcMNPV) factors involved in late gene expression (lefs), in the Sf9 insect cell-line, which is permissive for both viruses. It is well-established that nineteen AcMNPV lefs support optimal levels of activity from a late promoter-reporter gene cassette in this assay. A subgroup of SfMNPV lefs predicted to function in transcription-specific events substituted the corresponding AcMNPV lefs very efficiently. When all SfMNPV lefs were assayed, including replication lefs, activity was low, but addition of two AcMNPV lefs not encoded in SfMNPV genome, resulted in augmented reporter activity. SfMNPV IE-1 was able to activate an early promoter cis-linked to an hr-derived element from SfMNPV but not from AcMNPV. However, the level of early promoter activation with SfMNPV IE-1 was lower compared to AcMNPV IE-1.
我们利用瞬时表达分析,在对两种病毒均敏感的Sf9昆虫细胞系中,评估了杆状病毒草地贪夜蛾多角体病毒(SfMNPV)与苜蓿银纹夜蛾多角体病毒(AcMNPV)中参与晚期基因表达的因子(lef)的同源物的功能。众所周知,在该分析中,19个AcMNPV lef可支持晚期启动子 - 报告基因盒的最佳活性水平。预测在转录特异性事件中起作用的一组SfMNPV lef能够非常有效地替代相应的AcMNPV lef。当检测所有SfMNPV lef(包括复制lef)时,活性较低,但添加两个未在SfMNPV基因组中编码的AcMNPV lef后,报告基因活性增强。SfMNPV IE - 1能够激活与来自SfMNPV而非AcMNPV的hr衍生元件顺式连接的早期启动子。然而,与AcMNPV IE - 1相比,SfMNPV IE - 1对早期启动子的激活水平较低。