Cloning and Characterization of Sf9 Cell Lamin and the Lamin Conformational Changes during Autographa californica multiple nucleopolyhedrovirus Infection.
Cloning and Characterization of Sf9 Cell Lamin and the Lamin Conformational Changes during Autographa californica multiple nucleopolyhedrovirus Infection.
复制标题
Sf9细胞核纤层蛋白的克隆和表征以及苜蓿银纹夜蛾多重核多角体病毒感染过程中核纤层蛋白的构象变化
DOI：
10.3390/v8050126
复制
发表时间：
2016-05-07
期刊：
影响因子：
--
通讯作者：
Ji S
中科院分区：
文献类型：
--
作者：
Wei W;Wang H;Li X;Fang N;Yang S;Liu H;Kang X;Sun X;Ji S
目前，杆状病毒感染后板层改变的细节仍不清楚。本研究克隆了果翅夜蛾Sf9细胞系中的lamin基因。Sf9层蛋白的开放阅读框(ORF)为1860bp，编码一个相对分子质量为70 kDa的蛋白质。红色荧光蛋白(RFP)-层粘连蛋白融合蛋白的转染实验表明，Sf9层粘连蛋白定位于核缘。透射电子显微镜观察表明，褐斑潜蝇多核型多角体病毒(AcMNPV)核衣壳可穿透核膜。免疫荧光分析显示，在感染AcMNPV的Sf9细胞中，胞膜呈褶皱状染色，并形成内陷，而在模拟感染的Sf9细胞中，胞膜均匀分布在核周。Western blotting结果表明，杆状病毒感染的Sf9细胞的Lamin总量明显低于模型感染的细胞。这些结果表明，AcMNPV感染可诱导Sf9细胞膜结构和生化重排。
At present, the details of lamina alterations after baculovirus infection remain elusive. In this study, a lamin gene in the Sf9 cell line of Spodoptera frugiperda was cloned. The open reading frame (orf) of the Sf9 lamin was 1860 bp and encoded a protein with a molecular weight of 70 kDa. A transfection assay with a red fluorescence protein (rfp)-lamin fusion protein indicated that Sf9 lamin was localized in the nuclear rim. Transmission electron microscopy observations indicated that Autographa californica multiple nucleopolyhedrovirus (AcMNPV) nucleocapsids may pass through the nuclear envelope. Immunofluorescence assay indicated that the lamina showed a ruffled staining pattern with the formation of invaginations in the Sf9 cells infected with AcMNPV, while it was evenly distributed at the nuclear periphery of mock-infected cells. Western blotting results indicated that the total amount of lamin in the baculovirus-infected Sf9 cells was significantly decreased compared with the mock-infected cells. These results imply that AcMNPV infection induces structural and biochemical rearrangements of lamina of Sf9 cells.