Phenotypic grouping of 141 BmNPVs lacking viral gene sequences
Phenotypic grouping of 141 BmNPVs lacking viral gene sequences
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DOI：
10.1016/j.virusres.2012.02.016
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发表时间：
2012-05-01
期刊：
影响因子：
5
通讯作者：
Bando, Hisanori
中科院分区：
文献类型：
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作者：
Ono, Chikako;Kamagata, Takanori;Bando, Hisanori
我们使用BmNPVT3 bacmid系统（Ono et al., 2007）和lambda red重组系统（Datsenko and Wanner, 2000）构建了141个基因中每个基因的一系列基因敲除bmnpv (KOVs) （Gomi等，1999；Katsuma等，2011）。在随后使用插入多面蛋白位点的标记基因egfp（增强型绿色荧光蛋白基因）分析感染所需的特性时，将敲除病毒（kov）细分为a到d四种表型型，a型（86个kov）显示出与对照相当的扩展感染的能力，而b型（8个kov）传播感染的速度较慢。c型（37株kov）在转染的bmn细胞中表达egfp，但未观察到感染性病毒的产生。d型（10株）即使在转染实验中也没有表达egfp的能力。缺乏基因（pkip （Bm15）、gp41 （Bm66）、brod （Bm131）、Bm20、48、65、91、93或101）的kov被划分为可存活的a型和B型。(C) 2012 Elsevier B.V.版权所有
We constructed a series of gene knockout BmNPVs (KOVs) for each of 141 genes (Gomi et al., 1999; Katsuma et al., 2011)using the BmNPVT3 bacmid system (Ono et al., 2007) and lambda red recombination system (Datsenko and Wanner, 2000). In a subsequent analysis of the properties needed for infection using a marker gene, egfp (enhanced green fluorescent protein gene), inserted into the polyhedrin locus, the knockout viruses (KOVs) were subdivided into four phenotypic types, A to D. Type-A (86 KOVs) showed the ability to expand infections equivalent to the control while type-B (8 KOVs) spread infections more slowly. Type-C (37 KOVs) expressed egfp in transfected-BmN cells but the production of infectious viruses was not observed. Type-D (10 KOVs) showed no ability to express egfp even in the transfection experiments. KOVs lacking genes (pkip (Bm15), gp41 (Bm66), bro-d (Bm131), Bm20, 48, 65, 91, 93, or 101) previously identified as being essential, were placed in the viable type-A and B categories. (C) 2012 Elsevier B.V. All rights reserved.