Recombinant Bombyx mori Nucleopolyhedrovirus Harboring Green Fluorescent Protein

The gene encoding the green fluorescent protein (gfp) under the control of the highly expressed Autographa californica nucleopolyhedrovirus (AcMNPV)-polyhedrin promoter has been introduced into the polyhedron (polh) locus of Bombyx mori nucleopolyhedrovirus (BmNPV) by homologous recombination. The i...

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Main Authors: Deepak Sehgal, Karumathil P. Gopinathan
Format: Article
Language:English
Published: Taylor & Francis Group 1998-12-01
Series:BioTechniques
Online Access:https://www.future-science.com/doi/10.2144/98256st02
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author Deepak Sehgal
Karumathil P. Gopinathan
author_facet Deepak Sehgal
Karumathil P. Gopinathan
author_sort Deepak Sehgal
collection DOAJ
description The gene encoding the green fluorescent protein (gfp) under the control of the highly expressed Autographa californica nucleopolyhedrovirus (AcMNPV)-polyhedrin promoter has been introduced into the polyhedron (polh) locus of Bombyx mori nucleopolyhedrovirus (BmNPV) by homologous recombination. The insect host larvae and the cultured cells infected with this recombinant virus (vBmGFP) showed high levels of expression of gfp. The larval tissues permissive to virus multiplication could be readily visualized using the tagged recombinant virus, thus providing a direct approach to study the progress of virus infection or its control in the animal host. The highly expressed recombinant protein, GFP, could be easily solubilized from fat bodies. Thus, the caterpillar-based expression could serve as an economic alternative method for the large-scale production of recombinant proteins, even when they are nonsecretory in nature. Further, if the recombinant vBmGFP is used as a parent in generating other recombinants, conversion of the fluorescent plaques to colorless plaques serves as an easy means for screening recombinants. Such a method is especially helpful for BmNPV-recombinant selections in the absence of the other simplified techniques as are available for the prototype baculovirus AcMNPV system.
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spelling doaj-art-ca0dde5f50244e1ab11bdbbd5d84e2232025-08-20T02:25:37ZengTaylor & Francis GroupBioTechniques0736-62051940-98181998-12-01256997100610.2144/98256st02Recombinant Bombyx mori Nucleopolyhedrovirus Harboring Green Fluorescent ProteinDeepak Sehgal0Karumathil P. Gopinathan11Indian Institute of Science, Bangalore, India1Indian Institute of Science, Bangalore, IndiaThe gene encoding the green fluorescent protein (gfp) under the control of the highly expressed Autographa californica nucleopolyhedrovirus (AcMNPV)-polyhedrin promoter has been introduced into the polyhedron (polh) locus of Bombyx mori nucleopolyhedrovirus (BmNPV) by homologous recombination. The insect host larvae and the cultured cells infected with this recombinant virus (vBmGFP) showed high levels of expression of gfp. The larval tissues permissive to virus multiplication could be readily visualized using the tagged recombinant virus, thus providing a direct approach to study the progress of virus infection or its control in the animal host. The highly expressed recombinant protein, GFP, could be easily solubilized from fat bodies. Thus, the caterpillar-based expression could serve as an economic alternative method for the large-scale production of recombinant proteins, even when they are nonsecretory in nature. Further, if the recombinant vBmGFP is used as a parent in generating other recombinants, conversion of the fluorescent plaques to colorless plaques serves as an easy means for screening recombinants. Such a method is especially helpful for BmNPV-recombinant selections in the absence of the other simplified techniques as are available for the prototype baculovirus AcMNPV system.https://www.future-science.com/doi/10.2144/98256st02
spellingShingle Deepak Sehgal
Karumathil P. Gopinathan
Recombinant Bombyx mori Nucleopolyhedrovirus Harboring Green Fluorescent Protein
BioTechniques
title Recombinant Bombyx mori Nucleopolyhedrovirus Harboring Green Fluorescent Protein
title_full Recombinant Bombyx mori Nucleopolyhedrovirus Harboring Green Fluorescent Protein
title_fullStr Recombinant Bombyx mori Nucleopolyhedrovirus Harboring Green Fluorescent Protein
title_full_unstemmed Recombinant Bombyx mori Nucleopolyhedrovirus Harboring Green Fluorescent Protein
title_short Recombinant Bombyx mori Nucleopolyhedrovirus Harboring Green Fluorescent Protein
title_sort recombinant bombyx mori nucleopolyhedrovirus harboring green fluorescent protein
url https://www.future-science.com/doi/10.2144/98256st02
work_keys_str_mv AT deepaksehgal recombinantbombyxmorinucleopolyhedrovirusharboringgreenfluorescentprotein
AT karumathilpgopinathan recombinantbombyxmorinucleopolyhedrovirusharboringgreenfluorescentprotein