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Baculovirus entire ORF1629 is not essential for viral replication

It is generally accepted that ORF1629 is essential for baculovirus replication, which has enabled isolation of recombinant viruses in a baculovirus expression system using linearized viral DNA. ORF1629-defective viruses cannot replicate in insect cells; only recombinant virus with complete ORF1629 r...

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Autores principales: Gwak, Won Seok, Lee, See Nae, Choi, Jae Bang, Kim, Hyun Soo, Han, Beom Ku, Bae, Sung Min, Je, Yeon Ho, Woo, Soo Dong
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6706055/
https://www.ncbi.nlm.nih.gov/pubmed/31437242
http://dx.doi.org/10.1371/journal.pone.0221594
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author Gwak, Won Seok
Lee, See Nae
Choi, Jae Bang
Kim, Hyun Soo
Han, Beom Ku
Bae, Sung Min
Je, Yeon Ho
Woo, Soo Dong
author_facet Gwak, Won Seok
Lee, See Nae
Choi, Jae Bang
Kim, Hyun Soo
Han, Beom Ku
Bae, Sung Min
Je, Yeon Ho
Woo, Soo Dong
author_sort Gwak, Won Seok
collection PubMed
description It is generally accepted that ORF1629 is essential for baculovirus replication, which has enabled isolation of recombinant viruses in a baculovirus expression system using linearized viral DNA. ORF1629-defective viruses cannot replicate in insect cells; only recombinant virus with complete ORF1629 restoration by recombination can propagate, allowing for pure isolation and the development of bacmids for easy selection of recombinant viruses. We inadvertently found proliferation in insect cells of a bacmid lacking a complete ORF1629. PCR indicated no other viruses but a lack of complete ORF1629 in the proliferated bacmid, suggesting that the baculovirus propagated without a complete ORF1629. Lack of ORF1629 decreased the virus growth rate and yield; it also increased the occlusion body (OB) size but decreased its yield. These results were confirmed for Autographa californica multicapsid nucleopolyhedrovirus (AcMNPV) and Bombyx mori NPV (BmNPV). Thus, entire ORF1629 is not essential for viral replication, though it does affect the virus growth rate, yield, and size and OB production.
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spelling pubmed-67060552019-09-04 Baculovirus entire ORF1629 is not essential for viral replication Gwak, Won Seok Lee, See Nae Choi, Jae Bang Kim, Hyun Soo Han, Beom Ku Bae, Sung Min Je, Yeon Ho Woo, Soo Dong PLoS One Research Article It is generally accepted that ORF1629 is essential for baculovirus replication, which has enabled isolation of recombinant viruses in a baculovirus expression system using linearized viral DNA. ORF1629-defective viruses cannot replicate in insect cells; only recombinant virus with complete ORF1629 restoration by recombination can propagate, allowing for pure isolation and the development of bacmids for easy selection of recombinant viruses. We inadvertently found proliferation in insect cells of a bacmid lacking a complete ORF1629. PCR indicated no other viruses but a lack of complete ORF1629 in the proliferated bacmid, suggesting that the baculovirus propagated without a complete ORF1629. Lack of ORF1629 decreased the virus growth rate and yield; it also increased the occlusion body (OB) size but decreased its yield. These results were confirmed for Autographa californica multicapsid nucleopolyhedrovirus (AcMNPV) and Bombyx mori NPV (BmNPV). Thus, entire ORF1629 is not essential for viral replication, though it does affect the virus growth rate, yield, and size and OB production. Public Library of Science 2019-08-22 /pmc/articles/PMC6706055/ /pubmed/31437242 http://dx.doi.org/10.1371/journal.pone.0221594 Text en © 2019 Gwak et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Gwak, Won Seok
Lee, See Nae
Choi, Jae Bang
Kim, Hyun Soo
Han, Beom Ku
Bae, Sung Min
Je, Yeon Ho
Woo, Soo Dong
Baculovirus entire ORF1629 is not essential for viral replication
title Baculovirus entire ORF1629 is not essential for viral replication
title_full Baculovirus entire ORF1629 is not essential for viral replication
title_fullStr Baculovirus entire ORF1629 is not essential for viral replication
title_full_unstemmed Baculovirus entire ORF1629 is not essential for viral replication
title_short Baculovirus entire ORF1629 is not essential for viral replication
title_sort baculovirus entire orf1629 is not essential for viral replication
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6706055/
https://www.ncbi.nlm.nih.gov/pubmed/31437242
http://dx.doi.org/10.1371/journal.pone.0221594
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