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细胞依赖性的多角体产生和杆状病毒对 Autographa californica 多粒包埋核型多角体病毒 fp25k 突变体的在体和离体包埋。

Cell-dependent production of polyhedra and virion occlusion of Autographa californica multiple nucleopolyhedrovirus fp25k mutants in vitro and in vivo.

机构信息

Department of Microbiology, Miami University, Oxford, OH 45056, USA.

Graduate Program in Cell, Molecular and Structural Biology, Miami University, Oxford, OH 45056, USA.

出版信息

J Gen Virol. 2013 Jan;94(Pt 1):177-186. doi: 10.1099/vir.0.045591-0. Epub 2012 Sep 19.

Abstract

Members of the family Baculoviridae are insect-specific dsDNA viruses that have been used for biological control of insect pests in agriculture and forestry, as well as in research and pharmaceutical protein expression in insect cells and larvae. Autographa californica multiple nucleopolyhedrovirus (AcMNPV) is the type species of the family Baculoviridae. During infection of AcMNPV in permissive cells, fp25k mutants are positively selected, leading to the formation of the few polyhedra (FP) phenotype with reduced yield of polyhedra and reduced virion occlusion efficiency, which leads to decreased oral infectivity for insects. Here we report that polyhedra of AcMNPV fp25k mutants produced from different insect cell lines and insects have differences in larval per os infectivity, and that these variations are due to different virion occlusion efficiencies in these cell lines and insects. Polyhedra of AcMNPV fp25k mutants produced from Sf cells (Sf21 and Sf9, derived from Spodoptera frugiperda) and S. frugiperda larvae had poorer virion occlusion efficiency than those from Hi5 cells (derived from Trichoplusia ni) and T. ni larvae, based on immunoblots, DNA isolation and larval oral infection analysis. AcMNPV fp25k mutants formed clusters of FP and many polyhedra (MP) in the fat body cells of both T. ni and S. frugiperda larvae. Transmission electron microscopy revealed that the nature of virion occlusion of AcMNPV fp25k mutants was dependent on the different cells of the T. ni fat body tissue. Taken together, these results indicate that the FP phenotype and virion occlusion efficiency of fp25k mutants are influenced by the host insect cells.

摘要

杆状病毒科的成员是昆虫特异性双链 DNA 病毒,已被用于农业和林业中防治昆虫害虫,以及在昆虫细胞和幼虫中进行研究和制药蛋白表达。苜蓿银纹夜蛾多核型多角体病毒(AcMNPV)是杆状病毒科的模式种。在允许细胞中 AcMNPV 的感染过程中,fp25k 突变体被正向选择,导致形成少数多角体(FP)表型,多角体产量降低,病毒粒子包埋效率降低,从而降低了对昆虫的口服感染力。在这里,我们报告称,来自不同昆虫细胞系和昆虫的 AcMNPV fp25k 突变体产生的多角体在幼虫口服感染性方面存在差异,并且这些差异是由于这些细胞系和昆虫中的病毒粒子包埋效率不同所致。基于免疫印迹、DNA 分离和幼虫口服感染分析,来自 Sf 细胞(来自草地贪夜蛾的 Sf21 和 Sf9)和草地贪夜蛾幼虫的 AcMNPV fp25k 突变体的多角体的病毒粒子包埋效率低于来自 Hi5 细胞(来自烟青虫)和烟青虫幼虫的多角体。AcMNPV fp25k 突变体在两种烟青虫和草地贪夜蛾幼虫的脂肪体细胞中形成 FP 聚集体和许多多角体(MP)。透射电子显微镜显示,AcMNPV fp25k 突变体的病毒粒子包埋性质取决于烟青虫脂肪体组织的不同细胞。总之,这些结果表明,fp25k 突变体的 FP 表型和病毒粒子包埋效率受宿主昆虫细胞的影响。

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