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Cloning and Characterization of Sf9 Cell Lamin and the Lamin Conformational Changes during Autographa californica multiple nucleopolyhedrovirus Infection

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 transfe...

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Autores principales: Wei, Wenqiang, Wang, Hongju, Li, Xiaoya, Fang, Na, Yang, Shili, Liu, Hongyan, Kang, Xiaonan, Sun, Xiulian, Ji, Shaoping
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4885081/
https://www.ncbi.nlm.nih.gov/pubmed/27164127
http://dx.doi.org/10.3390/v8050126
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author Wei, Wenqiang
Wang, Hongju
Li, Xiaoya
Fang, Na
Yang, Shili
Liu, Hongyan
Kang, Xiaonan
Sun, Xiulian
Ji, Shaoping
author_facet Wei, Wenqiang
Wang, Hongju
Li, Xiaoya
Fang, Na
Yang, Shili
Liu, Hongyan
Kang, Xiaonan
Sun, Xiulian
Ji, Shaoping
author_sort Wei, Wenqiang
collection PubMed
description 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.
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spelling pubmed-48850812016-05-31 Cloning and Characterization of Sf9 Cell Lamin and the Lamin Conformational Changes during Autographa californica multiple nucleopolyhedrovirus Infection Wei, Wenqiang Wang, Hongju Li, Xiaoya Fang, Na Yang, Shili Liu, Hongyan Kang, Xiaonan Sun, Xiulian Ji, Shaoping Viruses Article 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. MDPI 2016-05-07 /pmc/articles/PMC4885081/ /pubmed/27164127 http://dx.doi.org/10.3390/v8050126 Text en © 2016 by the authors; licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC-BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Wei, Wenqiang
Wang, Hongju
Li, Xiaoya
Fang, Na
Yang, Shili
Liu, Hongyan
Kang, Xiaonan
Sun, Xiulian
Ji, Shaoping
Cloning and Characterization of Sf9 Cell Lamin and the Lamin Conformational Changes during Autographa californica multiple nucleopolyhedrovirus Infection
title Cloning and Characterization of Sf9 Cell Lamin and the Lamin Conformational Changes during Autographa californica multiple nucleopolyhedrovirus Infection
title_full Cloning and Characterization of Sf9 Cell Lamin and the Lamin Conformational Changes during Autographa californica multiple nucleopolyhedrovirus Infection
title_fullStr Cloning and Characterization of Sf9 Cell Lamin and the Lamin Conformational Changes during Autographa californica multiple nucleopolyhedrovirus Infection
title_full_unstemmed Cloning and Characterization of Sf9 Cell Lamin and the Lamin Conformational Changes during Autographa californica multiple nucleopolyhedrovirus Infection
title_short Cloning and Characterization of Sf9 Cell Lamin and the Lamin Conformational Changes during Autographa californica multiple nucleopolyhedrovirus Infection
title_sort cloning and characterization of sf9 cell lamin and the lamin conformational changes during autographa californica multiple nucleopolyhedrovirus infection
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4885081/
https://www.ncbi.nlm.nih.gov/pubmed/27164127
http://dx.doi.org/10.3390/v8050126
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