Roelvink P W, van Meer M M, de Kort C A, Possee R D, Hammock B D, Vlak J M
Department of Virology, Agricultural University Wageningen, The Netherlands.
J Gen Virol. 1992 Jun;73 ( Pt 6):1481-9. doi: 10.1099/0022-1317-73-6-1481.
The temporal expression of the Autographa californica multiple nucleocapsid nuclear polyhedrosis virus polyhedrin and p10 genes in Spodoptera frugiperda cells was studied using virus recombinants in which either gene was replaced by the juvenile hormone esterase (JHE) gene of Heliothis virescens. The JHE served as a highly specific and sensitive reporter for gene expression. Activation of the p10 gene followed a pattern different to that of polyhedrin. The p10 gene was activated a few hours earlier than the polyhedrin gene, but its expression reached a lower maximum level. Northern blot analysis complemented and confirmed the results obtained from the JHE assays. Co-infection of sense recombinants and those containing an antisense copy of the JHE gene in place of the polyhedrin or p10 gene resulted in reduced levels of JHE gene expression. These experiments independently supported the hypothesis that the p10 gene promoter is more active at earlier times post-infection than that of the polyhedrin gene. The results also highlight the potential of the antisense strategy as an experimental approach for the study of baculovirus gene regulation and possibly insect metabolism.
利用病毒重组体研究了苜蓿银纹夜蛾多核衣壳核型多角体病毒多角体蛋白基因和p10基因在草地贪夜蛾细胞中的时序表达,其中任一基因被棉铃虫的保幼激素酯酶(JHE)基因取代。JHE作为基因表达的高度特异性和灵敏性报告基因。p10基因的激活模式与多角体蛋白不同。p10基因比多角体蛋白基因早几个小时被激活,但其表达达到的最高水平较低。Northern印迹分析补充并证实了从JHE分析中获得的结果。有义重组体与含有取代多角体蛋白或p10基因的JHE基因反义拷贝的重组体共感染导致JHE基因表达水平降低。这些实验独立支持了以下假设:p10基因启动子在感染后早期比多角体蛋白基因启动子更活跃。结果还突出了反义策略作为研究杆状病毒基因调控以及可能的昆虫代谢的实验方法的潜力。