Development of a recombination system for the generation of occlusion positive genetically modified Anticarsia Gemmatalis Multiple Nucleopolyhedrovirus

Autores
Haase, Santiago; McCarthy, Christina Beryl; Ferrelli, María Leticia; Pidre, Matías Luis; Sciocco de Cap, Alicia; Romanowski, Víctor
Año de publicación
2015
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Anticarsia gemmatalis is an important pest in legume crops in South America and it has been successfully controlled using Anticarsia gemmatalis Multiple Nucleopolyhedrovirus (AgMNPV) in subtropical climate zones. Nevertheless, in temperate climates its speed of kill is too slow. Taking this into account, genetic modification of AgMNPV could lead to improvements of its biopesticidal properties. Here we report the generation of a two-component system that allows the production of recombinant AgMNPV. This system is based on a parental AgMNPV in which the polyhedrin gene (polh) was replaced by a bacterial β-galactosidase (lacZ) gene flanked by two target sites for the homing endonuclease I-PpoI. Co-transfection of insect cells with linearized (I-PpoI-digested) parental genome and a transfer vector allowed the restitution of polh and the expression of a heterologous gene upon homologous recombination, with a low background of non-recombinant AgMNPV. The system was validated by constructing a recombinant occlusion-positive (polh+) AgMNPV expressing the green fluorescent protein gene (gfp). This recombinant virus infected larvae normally per os and led to the expression of GFP in cell culture as well as in A. gemmatalis larvae. These results demonstrate that the system is an efficient method for the generation of recombinant AgMNPV expressing heterologous genes, which can be used for manifold purposes, including biotechnological and pharmaceutical applications and the production of orally infectious recombinants with improved biopesticidal properties.
Facultad de Ciencias Exactas
Instituto de Biotecnologia y Biologia Molecular
Materia
Ciencias Exactas
AgMNPV
Anticarsia gemmatalis
Bioinsecticide
Recombinant baculovirus
Velvetbean caterpillar
Nivel de accesibilidad
acceso abierto
Condiciones de uso
http://creativecommons.org/licenses/by/4.0/
Repositorio
SEDICI (UNLP)
Institución
Universidad Nacional de La Plata
OAI Identificador
oai:sedici.unlp.edu.ar:10915/85817
Acceder
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oai_identifier_str oai:sedici.unlp.edu.ar:10915/85817
network_acronym_str SEDICI
repository_id_str 1329
network_name_str SEDICI (UNLP)
spelling Development of a recombination system for the generation of occlusion positive genetically modified Anticarsia Gemmatalis Multiple NucleopolyhedrovirusHaase, SantiagoMcCarthy, Christina BerylFerrelli, María LeticiaPidre, Matías LuisSciocco de Cap, AliciaRomanowski, VíctorCiencias ExactasAgMNPVAnticarsia gemmatalisBioinsecticideRecombinant baculovirusVelvetbean caterpillar<i>Anticarsia gemmatalis</i> is an important pest in legume crops in South America and it has been successfully controlled using <i>Anticarsia gemmatalis Multiple Nucleopolyhedrovirus</i> (AgMNPV) in subtropical climate zones. Nevertheless, in temperate climates its speed of kill is too slow. Taking this into account, genetic modification of AgMNPV could lead to improvements of its biopesticidal properties. Here we report the generation of a two-component system that allows the production of recombinant AgMNPV. This system is based on a parental AgMNPV in which the polyhedrin gene (<i>polh</i>) was replaced by a bacterial β-<i>galactosidase</i> (<i>lacZ</i>) gene flanked by two target sites for the homing endonuclease I-<i>Ppo</i>I. Co-transfection of insect cells with linearized (I-<i>Ppo</i>I-digested) parental genome and a transfer vector allowed the restitution of <i>polh</i> and the expression of a heterologous gene upon homologous recombination, with a low background of non-recombinant AgMNPV. The system was validated by constructing a recombinant occlusion-positive (<i>polh</i><SUP>+</SUP>) AgMNPV expressing the green fluorescent protein gene (<i>gfp</i>). This recombinant virus infected larvae normally <i>per os</i> and led to the expression of GFP in cell culture as well as in <i>A. gemmatalis</i> larvae. These results demonstrate that the system is an efficient method for the generation of recombinant AgMNPV expressing heterologous genes, which can be used for manifold purposes, including biotechnological and pharmaceutical applications and the production of orally infectious recombinants with improved biopesticidal properties.Facultad de Ciencias ExactasInstituto de Biotecnologia y Biologia Molecular2015info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionArticulohttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdf1599-1612http://sedici.unlp.edu.ar/handle/10915/85817enginfo:eu-repo/semantics/altIdentifier/issn/1999-4915info:eu-repo/semantics/altIdentifier/doi/10.3390/v7041599info:eu-repo/semantics/openAccesshttp://creativecommons.org/licenses/by/4.0/Creative Commons Attribution 4.0 International (CC BY 4.0)reponame:SEDICI (UNLP)instname:Universidad Nacional de La Platainstacron:UNLP2025-09-10T12:19:38Zoai:sedici.unlp.edu.ar:10915/85817Institucionalhttp://sedici.unlp.edu.ar/Universidad públicaNo correspondehttp://sedici.unlp.edu.ar/oai/snrdalira@sedici.unlp.edu.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:13292025-09-10 12:19:38.503SEDICI (UNLP) - Universidad Nacional de La Platafalse
dc.title.none.fl_str_mv Development of a recombination system for the generation of occlusion positive genetically modified Anticarsia Gemmatalis Multiple Nucleopolyhedrovirus
title Development of a recombination system for the generation of occlusion positive genetically modified Anticarsia Gemmatalis Multiple Nucleopolyhedrovirus
spellingShingle Development of a recombination system for the generation of occlusion positive genetically modified Anticarsia Gemmatalis Multiple Nucleopolyhedrovirus
Haase, Santiago
Ciencias Exactas
AgMNPV
Anticarsia gemmatalis
Bioinsecticide
Recombinant baculovirus
Velvetbean caterpillar
title_short Development of a recombination system for the generation of occlusion positive genetically modified Anticarsia Gemmatalis Multiple Nucleopolyhedrovirus
title_full Development of a recombination system for the generation of occlusion positive genetically modified Anticarsia Gemmatalis Multiple Nucleopolyhedrovirus
title_fullStr Development of a recombination system for the generation of occlusion positive genetically modified Anticarsia Gemmatalis Multiple Nucleopolyhedrovirus
title_full_unstemmed Development of a recombination system for the generation of occlusion positive genetically modified Anticarsia Gemmatalis Multiple Nucleopolyhedrovirus
title_sort Development of a recombination system for the generation of occlusion positive genetically modified Anticarsia Gemmatalis Multiple Nucleopolyhedrovirus
dc.creator.none.fl_str_mv Haase, Santiago
McCarthy, Christina Beryl
Ferrelli, María Leticia
Pidre, Matías Luis
Sciocco de Cap, Alicia
Romanowski, Víctor
author Haase, Santiago
author_facet Haase, Santiago
McCarthy, Christina Beryl
Ferrelli, María Leticia
Pidre, Matías Luis
Sciocco de Cap, Alicia
Romanowski, Víctor
author_role author
author2 McCarthy, Christina Beryl
Ferrelli, María Leticia
Pidre, Matías Luis
Sciocco de Cap, Alicia
Romanowski, Víctor
author2_role author
author
author
author
author
dc.subject.none.fl_str_mv Ciencias Exactas
AgMNPV
Anticarsia gemmatalis
Bioinsecticide
Recombinant baculovirus
Velvetbean caterpillar
topic Ciencias Exactas
AgMNPV
Anticarsia gemmatalis
Bioinsecticide
Recombinant baculovirus
Velvetbean caterpillar
dc.description.none.fl_txt_mv <i>Anticarsia gemmatalis</i> is an important pest in legume crops in South America and it has been successfully controlled using <i>Anticarsia gemmatalis Multiple Nucleopolyhedrovirus</i> (AgMNPV) in subtropical climate zones. Nevertheless, in temperate climates its speed of kill is too slow. Taking this into account, genetic modification of AgMNPV could lead to improvements of its biopesticidal properties. Here we report the generation of a two-component system that allows the production of recombinant AgMNPV. This system is based on a parental AgMNPV in which the polyhedrin gene (<i>polh</i>) was replaced by a bacterial β-<i>galactosidase</i> (<i>lacZ</i>) gene flanked by two target sites for the homing endonuclease I-<i>Ppo</i>I. Co-transfection of insect cells with linearized (I-<i>Ppo</i>I-digested) parental genome and a transfer vector allowed the restitution of <i>polh</i> and the expression of a heterologous gene upon homologous recombination, with a low background of non-recombinant AgMNPV. The system was validated by constructing a recombinant occlusion-positive (<i>polh</i><SUP>+</SUP>) AgMNPV expressing the green fluorescent protein gene (<i>gfp</i>). This recombinant virus infected larvae normally <i>per os</i> and led to the expression of GFP in cell culture as well as in <i>A. gemmatalis</i> larvae. These results demonstrate that the system is an efficient method for the generation of recombinant AgMNPV expressing heterologous genes, which can be used for manifold purposes, including biotechnological and pharmaceutical applications and the production of orally infectious recombinants with improved biopesticidal properties.
Facultad de Ciencias Exactas
Instituto de Biotecnologia y Biologia Molecular
description <i>Anticarsia gemmatalis</i> is an important pest in legume crops in South America and it has been successfully controlled using <i>Anticarsia gemmatalis Multiple Nucleopolyhedrovirus</i> (AgMNPV) in subtropical climate zones. Nevertheless, in temperate climates its speed of kill is too slow. Taking this into account, genetic modification of AgMNPV could lead to improvements of its biopesticidal properties. Here we report the generation of a two-component system that allows the production of recombinant AgMNPV. This system is based on a parental AgMNPV in which the polyhedrin gene (<i>polh</i>) was replaced by a bacterial β-<i>galactosidase</i> (<i>lacZ</i>) gene flanked by two target sites for the homing endonuclease I-<i>Ppo</i>I. Co-transfection of insect cells with linearized (I-<i>Ppo</i>I-digested) parental genome and a transfer vector allowed the restitution of <i>polh</i> and the expression of a heterologous gene upon homologous recombination, with a low background of non-recombinant AgMNPV. The system was validated by constructing a recombinant occlusion-positive (<i>polh</i><SUP>+</SUP>) AgMNPV expressing the green fluorescent protein gene (<i>gfp</i>). This recombinant virus infected larvae normally <i>per os</i> and led to the expression of GFP in cell culture as well as in <i>A. gemmatalis</i> larvae. These results demonstrate that the system is an efficient method for the generation of recombinant AgMNPV expressing heterologous genes, which can be used for manifold purposes, including biotechnological and pharmaceutical applications and the production of orally infectious recombinants with improved biopesticidal properties.
publishDate 2015
dc.date.none.fl_str_mv 2015
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
Articulo
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://sedici.unlp.edu.ar/handle/10915/85817
url http://sedici.unlp.edu.ar/handle/10915/85817
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/issn/1999-4915
info:eu-repo/semantics/altIdentifier/doi/10.3390/v7041599
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by/4.0/
Creative Commons Attribution 4.0 International (CC BY 4.0)
eu_rights_str_mv openAccess
rights_invalid_str_mv http://creativecommons.org/licenses/by/4.0/
Creative Commons Attribution 4.0 International (CC BY 4.0)
dc.format.none.fl_str_mv application/pdf
1599-1612
dc.source.none.fl_str_mv reponame:SEDICI (UNLP)
instname:Universidad Nacional de La Plata
instacron:UNLP
reponame_str SEDICI (UNLP)
collection SEDICI (UNLP)
instname_str Universidad Nacional de La Plata
instacron_str UNLP
institution UNLP
repository.name.fl_str_mv SEDICI (UNLP) - Universidad Nacional de La Plata
repository.mail.fl_str_mv alira@sedici.unlp.edu.ar
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score 12.993085

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