Baculovirus expression of cloned porcine arterivirus generates infectious particles in both insect and mammalian cells

Studies on several viral pathogens have been hampered by the lack of appropriate in vitro systems for their propagation and amplification. Porcine reproductive and respiratory syndrome virus (PRRSV), an arterivirus containing a single-stranded positive-sense RNA genome (∼15 kb), was served as a mode...

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Autores principales: Zheng, Hao, Liu, Changlong, Zhuang, Jinshan, Yuan, Shishan
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier B.V. 2010
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7114269/
https://www.ncbi.nlm.nih.gov/pubmed/20728481
http://dx.doi.org/10.1016/j.jbiotec.2010.08.009
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author Zheng, Hao
Liu, Changlong
Zhuang, Jinshan
Yuan, Shishan
author_facet Zheng, Hao
Liu, Changlong
Zhuang, Jinshan
Yuan, Shishan
author_sort Zheng, Hao
collection PubMed
description Studies on several viral pathogens have been hampered by the lack of appropriate in vitro systems for their propagation and amplification. Porcine reproductive and respiratory syndrome virus (PRRSV), an arterivirus containing a single-stranded positive-sense RNA genome (∼15 kb), was served as a model virus and its genomic cDNA was recombinated into baculovirus. We investigated whether infectious virus particles could be generated by expression of the full-length cloned genome from the modified baculovirus vector. The recombinant baculovirus, AcAPRRS, was used to infect sf9 cells. Immunofluorescence assay demonstrated the presence of PRRSV nonstructural protein (nsp) 2 and nucleocapsid (N) protein and electron microscopy revealed PRRSV particles in the culture supernatant. Infectious PRRSV particles were also produced in susceptible MARC-145 cells inoculated with AcAPRRS, and the growth characteristics of the PRRSV generated were similar to those of the parental PRRSV strain. Infectious PRRSV particles were also generated following AcAPRRS transduction of BHK-21 cells and Vero cells that are not sensitive to PRRSV. Titers of PRRSV obtained from BHK-21 and Vero cells were up to 10(4.05) TCID(50)/ml. These findings open a new route to the propagation of the virus in vitro and will be of utility in vaccine development.
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spelling pubmed-71142692020-04-02 Baculovirus expression of cloned porcine arterivirus generates infectious particles in both insect and mammalian cells Zheng, Hao Liu, Changlong Zhuang, Jinshan Yuan, Shishan J Biotechnol Article Studies on several viral pathogens have been hampered by the lack of appropriate in vitro systems for their propagation and amplification. Porcine reproductive and respiratory syndrome virus (PRRSV), an arterivirus containing a single-stranded positive-sense RNA genome (∼15 kb), was served as a model virus and its genomic cDNA was recombinated into baculovirus. We investigated whether infectious virus particles could be generated by expression of the full-length cloned genome from the modified baculovirus vector. The recombinant baculovirus, AcAPRRS, was used to infect sf9 cells. Immunofluorescence assay demonstrated the presence of PRRSV nonstructural protein (nsp) 2 and nucleocapsid (N) protein and electron microscopy revealed PRRSV particles in the culture supernatant. Infectious PRRSV particles were also produced in susceptible MARC-145 cells inoculated with AcAPRRS, and the growth characteristics of the PRRSV generated were similar to those of the parental PRRSV strain. Infectious PRRSV particles were also generated following AcAPRRS transduction of BHK-21 cells and Vero cells that are not sensitive to PRRSV. Titers of PRRSV obtained from BHK-21 and Vero cells were up to 10(4.05) TCID(50)/ml. These findings open a new route to the propagation of the virus in vitro and will be of utility in vaccine development. Elsevier B.V. 2010-10-15 2010-08-20 /pmc/articles/PMC7114269/ /pubmed/20728481 http://dx.doi.org/10.1016/j.jbiotec.2010.08.009 Text en Copyright © 2010 Elsevier B.V. All rights reserved. Since January 2020 Elsevier has created a COVID-19 resource centre with free information in English and Mandarin on the novel coronavirus COVID-19. The COVID-19 resource centre is hosted on Elsevier Connect, the company's public news and information website. Elsevier hereby grants permission to make all its COVID-19-related research that is available on the COVID-19 resource centre - including this research content - immediately available in PubMed Central and other publicly funded repositories, such as the WHO COVID database with rights for unrestricted research re-use and analyses in any form or by any means with acknowledgement of the original source. These permissions are granted for free by Elsevier for as long as the COVID-19 resource centre remains active.
spellingShingle Article
Zheng, Hao
Liu, Changlong
Zhuang, Jinshan
Yuan, Shishan
Baculovirus expression of cloned porcine arterivirus generates infectious particles in both insect and mammalian cells
title Baculovirus expression of cloned porcine arterivirus generates infectious particles in both insect and mammalian cells
title_full Baculovirus expression of cloned porcine arterivirus generates infectious particles in both insect and mammalian cells
title_fullStr Baculovirus expression of cloned porcine arterivirus generates infectious particles in both insect and mammalian cells
title_full_unstemmed Baculovirus expression of cloned porcine arterivirus generates infectious particles in both insect and mammalian cells
title_short Baculovirus expression of cloned porcine arterivirus generates infectious particles in both insect and mammalian cells
title_sort baculovirus expression of cloned porcine arterivirus generates infectious particles in both insect and mammalian cells
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7114269/
https://www.ncbi.nlm.nih.gov/pubmed/20728481
http://dx.doi.org/10.1016/j.jbiotec.2010.08.009
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