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

<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...

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Autores principales: Haase, Santiago, McCarthy, Christina Beryl, Ferrelli, María Leticia, Pidre, Matías Luis, Sciocco de Cap, Alicia, Romanowski, Víctor
Formato: Articulo
Lenguaje:Inglés
Publicado: 2015
Materias:
Ciencias Exactas
AgMNPV
Anticarsia gemmatalis
Bioinsecticide
Recombinant baculovirus
Velvetbean caterpillar
Acceso en línea:http://sedici.unlp.edu.ar/handle/10915/85817
Aporte de:
SEDICI (UNLP) de Universidad Nacional de La Plata
id I19-R120-10915-85817
record_format dspace
institution Universidad Nacional de La Plata
institution_str I-19
repository_str R-120
collection SEDICI (UNLP)
language Inglés
topic Ciencias Exactas
AgMNPV
Anticarsia gemmatalis
Bioinsecticide
Recombinant baculovirus
Velvetbean caterpillar
spellingShingle Ciencias Exactas
AgMNPV
Anticarsia gemmatalis
Bioinsecticide
Recombinant baculovirus
Velvetbean caterpillar
Haase, Santiago
McCarthy, Christina Beryl
Ferrelli, María Leticia
Pidre, Matías Luis
Sciocco de Cap, Alicia
Romanowski, Víctor
Development of a recombination system for the generation of occlusion positive genetically modified Anticarsia Gemmatalis Multiple Nucleopolyhedrovirus
topic_facet Ciencias Exactas
AgMNPV
Anticarsia gemmatalis
Bioinsecticide
Recombinant baculovirus
Velvetbean caterpillar
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.
format Articulo
Articulo
author Haase, Santiago
McCarthy, Christina Beryl
Ferrelli, María Leticia
Pidre, Matías Luis
Sciocco de Cap, Alicia
Romanowski, Víctor
author_facet Haase, Santiago
McCarthy, Christina Beryl
Ferrelli, María Leticia
Pidre, Matías Luis
Sciocco de Cap, Alicia
Romanowski, Víctor
author_sort Haase, Santiago
title Development of a recombination system for the generation of occlusion positive genetically modified Anticarsia Gemmatalis Multiple Nucleopolyhedrovirus
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
publishDate 2015
url http://sedici.unlp.edu.ar/handle/10915/85817
work_keys_str_mv AT haasesantiago developmentofarecombinationsystemforthegenerationofocclusionpositivegeneticallymodifiedanticarsiagemmatalismultiplenucleopolyhedrovirus
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AT ferrellimarialeticia developmentofarecombinationsystemforthegenerationofocclusionpositivegeneticallymodifiedanticarsiagemmatalismultiplenucleopolyhedrovirus
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bdutipo_str Repositorios
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