CHARACTERIZATION OF PRODUCTIVE AND NONPRODUCTIVE ACMNPV INFECTION IN SELECTED INSECT-CELL LINES
CHARACTERIZATION OF PRODUCTIVE AND NONPRODUCTIVE ACMNPV INFECTION IN SELECTED INSECT-CELL LINES
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DOI：
10.1006/viro.1993.1595
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发表时间：
1993-11-01
期刊：
影响因子：
3.7
通讯作者：
MILLER, LK
中科院分区：
文献类型：
--
作者：
MORRIS, TD;MILLER, LK
我们研究了AcMNPV在多种培养的昆虫细胞中的生产性和非生产性感染过程，这些昆虫细胞包括来自草地贪夜蛾（IPLB-SF-21）、甘蓝夜蛾（SES-MaBr-3）、烟羽夜蛾（IPRL-CF-1）、家蚕（BmN-4）、舞毒蛾（IPLB-Ld 652 Y）、玉米夜蛾（Hz 1b 3）和黑腹果蝇（Dm）的品系。在每个细胞系中，我们已经检查了早期，晚期和非常晚期启动子的病毒利用，病毒DNA的复制和出芽病毒（BV）和多面体包涵体（PIB）的产生。使用荧光激活细胞分选（FACS）技术在单细胞水平上进行启动子使用的分析。细胞培养物用含有CAT报告基因的重组病毒感染，所述CAT报告基因在病毒早期、晚期或极晚期启动子的转录控制下。对细胞进行免疫染色以检测CAT基因产物，并通过FACS分析确定基于每个细胞的感染细胞的相对CAT含量。IPLB-SF 21细胞的生产性感染基本上涉及100%的细胞，而对生产性较低的细胞系（IPRL-CF-1和SES-MaBr-3）的分析表明，病毒仅在细胞亚群中启动和完成复制过程。对非生产性感染（在BmN-4、IPLB-Ld 652 Y、Hz 1b 3、DrosophilaSchneider中）的分析揭示了在每种细胞系中病毒DNA复制和启动子使用的不同模式，这表明生产性感染的各种障碍。
We have investigated the process of productive and non-productive AcMNPV infection in a variety of cultured insect cells including lines derived fromSpodoptera frugiperda(IPLB-SF21),Mamestra brassicae(SES-MaBr-3),Choristoneura fumiferana(IPRL-CF-1),Bombyx mori(Bm N-4),Lymantria dispar(IPLB-Ld652Y),Helicoverpa zea(Hz1 b3), andDrosophila melanogaster(Dm). In each cell line, we have examined viral utilization of an early, a late, and a very late promoter, the replication of viral DNA and the production of budded virus (BV) and polyhedral inclusion bodies (PIBs). Analysis of promoter use on the single cell level was performed using fluorescence-activated cell sorting (FACS) technology. Cell cultures were infected with recombinant viruses containing CAT reporter genes under the transcriptional control of viral early, late or very late promoters. Cells were immunostained to detect the CAT gene product and the relative CAT content of infected cells on a per cell basis was determined by FACS analysis. Productive infection of IPLB-SF21 cells involved essentially 100% of the cells while analysis of less productive lines (IPRL-CF-1 and SES-MaBr-3) suggests that the virus initiates and completes the replication process only in a subpopulation of cells. Analysis of non-productive infections (in BmN-4, IPLB-Ld652Y, Hz1b3,DrosophilaSchneider) revealed different patterns of viral DNA replication and promoter use in each cell line suggesting a variety of obstacles to productive infection.