Tian T, Medina V, Mayhew D E, Maeda S, Falk B W
Department of Plant Pathology, University of California, Davis 95616, USA.
Virology. 1995 Oct 20;213(1):204-12. doi: 10.1006/viro.1995.1560.
We subcloned various constructs of the beet western yellows luteovirus (BWYV) 3' proximal genes into Bombyx mori nuclear polyhedrosis virus (BmNPV) transfer vectors and constructed recombinant BmNPVs. The recombinant BmNPVs were used to infect BmN cells and B. mori larvae. Protein expression was analyzed by SDS-PAGE and immunoblot analysis, and the BWYV-encoded capsid protein (CP), P19, readthrough protein (RT), and a modified capsid-readthrough protein (CP-RT*) were detected in BmN cells infected by the respective BmNPVs. However, we detected only the BWYV CP and P19 in BmN cells infected with a recombinant BmNPV containing all three BWYV 3' proximal genes. BmN cells and fat body cells from B. mori larvae infected with recombinant BmNPVs were analyzed by transmission electron microscopy and immunogold labeling. Particles which morphologically and serologically resembled BWYV virions were detected in the nuclei of BmN cells and B. mori larval fat body cells infected with recombinant BmNPVs producing either the BWYV CP alone or BWYV CP plus a BWYV CP-RT*. Similar particles were not detected in cells infected with a control BmNPV or recombinant BmNPV producing only CP-RT*.
我们将甜菜西方黄化黄症病毒(BWYV)3'近端基因的各种构建体亚克隆到家蚕核型多角体病毒(BmNPV)转移载体中,并构建了重组BmNPV。这些重组BmNPV用于感染BmN细胞和家蚕幼虫。通过SDS-PAGE和免疫印迹分析对蛋白质表达进行分析,在被相应BmNPV感染的BmN细胞中检测到了BWYV编码的衣壳蛋白(CP)、P19、通读蛋白(RT)以及一种修饰的衣壳-通读蛋白(CP-RT*)。然而,在感染了包含所有三个BWYV 3'近端基因的重组BmNPV的BmN细胞中,我们仅检测到了BWYV CP和P19。通过透射电子显微镜和免疫金标记对感染重组BmNPV的家蚕幼虫的BmN细胞和脂肪体细胞进行分析。在感染了单独产生BWYV CP或产生BWYV CP加BWYV CP-RT的重组BmNPV的BmN细胞和家蚕幼虫脂肪体细胞的细胞核中,检测到了形态和血清学上类似于BWYV病毒粒子的颗粒。在感染对照BmNPV或仅产生CP-RT的重组BmNPV的细胞中未检测到类似颗粒。