Olszewski Julie A, Dall David J
CSIRO Entomology, GPO Box 1700, Canberra ACT 2601, Australia1.
J Gen Virol. 2002 Feb;83(Pt 2):451-461. doi: 10.1099/0022-1317-83-2-451.
This report describes the first production of recombinant forms of Heliothis (Helicoverpa) armigera entomopoxvirus (HaEPV). These HaEPVs are engineered at either the spheroidin or fusolin locus, to produce the green fluorescent marker protein (GFP). The growth properties of these recombinant HaEPVs, in comparison to the parental HaEPV, were assessed in cultured Spodoptera frugiperda Sf9 cells. Additionally, GFP production by these recombinant HaEPVs was compared to that of a GFP-expressing recombinant of the baculovirus Autographa californica nucleopolyhedrovirus (AcNPV) in the same in vitro system, at various multiplicities of infection. Expression of GFP from the HaEPV spheroidin locus produced up to 60% of that generated from the AcNPV polyhedrin locus, albeit over a longer period of infection. A considerably lower yield was recorded from the HaEPV fusolin locus, a result that contrasted markedly with the apparent activity of this promoter in caterpillar infections in vivo. The potential applications for further development of HaEPV expression systems are discussed.
本报告描述了棉铃虫昆虫痘病毒(HaEPV)重组形式的首次产生。这些HaEPV在多角体蛋白或融合素基因座处进行改造,以产生绿色荧光标记蛋白(GFP)。在培养的草地贪夜蛾Sf9细胞中评估了这些重组HaEPV与亲本HaEPV相比的生长特性。此外,在相同的体外系统中,以不同的感染复数,将这些重组HaEPV产生GFP的情况与杆状病毒苜蓿银纹夜蛾核多角体病毒(AcNPV)的GFP表达重组体进行了比较。来自HaEPV多角体蛋白基因座的GFP表达量高达AcNPV多角体蛋白基因座产生量的60%,尽管感染时间更长。从HaEPV融合素基因座记录到的产量要低得多,这一结果与该启动子在体内毛虫感染中的明显活性形成了鲜明对比。讨论了HaEPV表达系统进一步开发的潜在应用。