Trans-complementation of polyhedrin by a stably transformed Sf9 insect cell line allows occ- baculovirus occlusion and larval per os infectivity

Autores
López, María Gabriela; Alfonso, Victoria; Carrillo, Elisa Cristina; Taboga, Oscar Alberto
Año de publicación
2010
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Oral infection of insect larvae with baculovirus is an advantageous methodology for producing high levels of recombinant proteins and for achieving plague control. However, many recombinant baculoviruses express a foreign protein in lieu of the polyhedrin and hence do not form occlusion bodies (occ-), resulting in extremely reduced per os infectivity in larvae. To overcome this limitation, stably transformed insect cell lines expressing polyhedrin capable of occluding occ- recombinant baculovirus by trans-complementation were developed to obtain oral inoculum for insect larvae infection. First, the optimum regulatory region of polyhedrin promoter was determined utilizing chloramphenicol acetyl transferase (CAT) as the reporter gene. After infection with occ- baculovirus, the higher expression levels of CAT were achieved when a region of 2735 bp that contained sequences known to have transcriptional enhancer functions were present upstream the polyhedrin coding sequence. This regulatory region was selected to drive polyhedrin expression in insect cell lines. Transfection of Sf9 cells with plasmid carrying polyhedrin gene and stable cell lines established by selection with blasticidin showed polyhedrin expression and, moreover, crystalline polyhedron-like structures were visualized by optic microscopy. Oral infectivity was demonstrated by fluorescence detection in Rachiplusia nu larvae infected with occluded AcGFPpolh- baculovirus obtained using the system presented here. © 2009 Elsevier B.V. All rights reserved.
Fil: López, María Gabriela. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Alfonso, Victoria. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Carrillo, Elisa Cristina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas; Argentina
Fil: Taboga, Oscar Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas; Argentina
Materia
Baculovirus
Gfp
Polyhedrin
Sf9
Stable Cell Lines
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/53645
Acceder
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network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Trans-complementation of polyhedrin by a stably transformed Sf9 insect cell line allows occ- baculovirus occlusion and larval per os infectivityLópez, María GabrielaAlfonso, VictoriaCarrillo, Elisa CristinaTaboga, Oscar AlbertoBaculovirusGfpPolyhedrinSf9Stable Cell Lineshttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Oral infection of insect larvae with baculovirus is an advantageous methodology for producing high levels of recombinant proteins and for achieving plague control. However, many recombinant baculoviruses express a foreign protein in lieu of the polyhedrin and hence do not form occlusion bodies (occ-), resulting in extremely reduced per os infectivity in larvae. To overcome this limitation, stably transformed insect cell lines expressing polyhedrin capable of occluding occ- recombinant baculovirus by trans-complementation were developed to obtain oral inoculum for insect larvae infection. First, the optimum regulatory region of polyhedrin promoter was determined utilizing chloramphenicol acetyl transferase (CAT) as the reporter gene. After infection with occ- baculovirus, the higher expression levels of CAT were achieved when a region of 2735 bp that contained sequences known to have transcriptional enhancer functions were present upstream the polyhedrin coding sequence. This regulatory region was selected to drive polyhedrin expression in insect cell lines. Transfection of Sf9 cells with plasmid carrying polyhedrin gene and stable cell lines established by selection with blasticidin showed polyhedrin expression and, moreover, crystalline polyhedron-like structures were visualized by optic microscopy. Oral infectivity was demonstrated by fluorescence detection in Rachiplusia nu larvae infected with occluded AcGFPpolh- baculovirus obtained using the system presented here. © 2009 Elsevier B.V. All rights reserved.Fil: López, María Gabriela. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Alfonso, Victoria. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Carrillo, Elisa Cristina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas; ArgentinaFil: Taboga, Oscar Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas; ArgentinaElsevier Science2010-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/53645López, María Gabriela; Alfonso, Victoria; Carrillo, Elisa Cristina; Taboga, Oscar Alberto; Trans-complementation of polyhedrin by a stably transformed Sf9 insect cell line allows occ- baculovirus occlusion and larval per os infectivity; Elsevier Science; Journal of Biotechnology; 145; 2; 1-2010; 199-2050168-1656CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.1016/j.jbiotec.2009.10.015info:eu-repo/semantics/altIdentifier/url/https://www.sciencedirect.com/science/article/pii/S0168165609004842info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-03T10:06:24Zoai:ri.conicet.gov.ar:11336/53645instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-09-03 10:06:24.465CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Trans-complementation of polyhedrin by a stably transformed Sf9 insect cell line allows occ- baculovirus occlusion and larval per os infectivity
title Trans-complementation of polyhedrin by a stably transformed Sf9 insect cell line allows occ- baculovirus occlusion and larval per os infectivity
spellingShingle Trans-complementation of polyhedrin by a stably transformed Sf9 insect cell line allows occ- baculovirus occlusion and larval per os infectivity
López, María Gabriela
Baculovirus
Gfp
Polyhedrin
Sf9
Stable Cell Lines
title_short Trans-complementation of polyhedrin by a stably transformed Sf9 insect cell line allows occ- baculovirus occlusion and larval per os infectivity
title_full Trans-complementation of polyhedrin by a stably transformed Sf9 insect cell line allows occ- baculovirus occlusion and larval per os infectivity
title_fullStr Trans-complementation of polyhedrin by a stably transformed Sf9 insect cell line allows occ- baculovirus occlusion and larval per os infectivity
title_full_unstemmed Trans-complementation of polyhedrin by a stably transformed Sf9 insect cell line allows occ- baculovirus occlusion and larval per os infectivity
title_sort Trans-complementation of polyhedrin by a stably transformed Sf9 insect cell line allows occ- baculovirus occlusion and larval per os infectivity
dc.creator.none.fl_str_mv López, María Gabriela
Alfonso, Victoria
Carrillo, Elisa Cristina
Taboga, Oscar Alberto
author López, María Gabriela
author_facet López, María Gabriela
Alfonso, Victoria
Carrillo, Elisa Cristina
Taboga, Oscar Alberto
author_role author
author2 Alfonso, Victoria
Carrillo, Elisa Cristina
Taboga, Oscar Alberto
author2_role author
author
author
dc.subject.none.fl_str_mv Baculovirus
Gfp
Polyhedrin
Sf9
Stable Cell Lines
topic Baculovirus
Gfp
Polyhedrin
Sf9
Stable Cell Lines
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Oral infection of insect larvae with baculovirus is an advantageous methodology for producing high levels of recombinant proteins and for achieving plague control. However, many recombinant baculoviruses express a foreign protein in lieu of the polyhedrin and hence do not form occlusion bodies (occ-), resulting in extremely reduced per os infectivity in larvae. To overcome this limitation, stably transformed insect cell lines expressing polyhedrin capable of occluding occ- recombinant baculovirus by trans-complementation were developed to obtain oral inoculum for insect larvae infection. First, the optimum regulatory region of polyhedrin promoter was determined utilizing chloramphenicol acetyl transferase (CAT) as the reporter gene. After infection with occ- baculovirus, the higher expression levels of CAT were achieved when a region of 2735 bp that contained sequences known to have transcriptional enhancer functions were present upstream the polyhedrin coding sequence. This regulatory region was selected to drive polyhedrin expression in insect cell lines. Transfection of Sf9 cells with plasmid carrying polyhedrin gene and stable cell lines established by selection with blasticidin showed polyhedrin expression and, moreover, crystalline polyhedron-like structures were visualized by optic microscopy. Oral infectivity was demonstrated by fluorescence detection in Rachiplusia nu larvae infected with occluded AcGFPpolh- baculovirus obtained using the system presented here. © 2009 Elsevier B.V. All rights reserved.
Fil: López, María Gabriela. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Alfonso, Victoria. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Carrillo, Elisa Cristina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas; Argentina
Fil: Taboga, Oscar Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas; Argentina
description Oral infection of insect larvae with baculovirus is an advantageous methodology for producing high levels of recombinant proteins and for achieving plague control. However, many recombinant baculoviruses express a foreign protein in lieu of the polyhedrin and hence do not form occlusion bodies (occ-), resulting in extremely reduced per os infectivity in larvae. To overcome this limitation, stably transformed insect cell lines expressing polyhedrin capable of occluding occ- recombinant baculovirus by trans-complementation were developed to obtain oral inoculum for insect larvae infection. First, the optimum regulatory region of polyhedrin promoter was determined utilizing chloramphenicol acetyl transferase (CAT) as the reporter gene. After infection with occ- baculovirus, the higher expression levels of CAT were achieved when a region of 2735 bp that contained sequences known to have transcriptional enhancer functions were present upstream the polyhedrin coding sequence. This regulatory region was selected to drive polyhedrin expression in insect cell lines. Transfection of Sf9 cells with plasmid carrying polyhedrin gene and stable cell lines established by selection with blasticidin showed polyhedrin expression and, moreover, crystalline polyhedron-like structures were visualized by optic microscopy. Oral infectivity was demonstrated by fluorescence detection in Rachiplusia nu larvae infected with occluded AcGFPpolh- baculovirus obtained using the system presented here. © 2009 Elsevier B.V. All rights reserved.
publishDate 2010
dc.date.none.fl_str_mv 2010-01
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
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://hdl.handle.net/11336/53645
López, María Gabriela; Alfonso, Victoria; Carrillo, Elisa Cristina; Taboga, Oscar Alberto; Trans-complementation of polyhedrin by a stably transformed Sf9 insect cell line allows occ- baculovirus occlusion and larval per os infectivity; Elsevier Science; Journal of Biotechnology; 145; 2; 1-2010; 199-205
0168-1656
CONICET Digital
CONICET
url http://hdl.handle.net/11336/53645
identifier_str_mv López, María Gabriela; Alfonso, Victoria; Carrillo, Elisa Cristina; Taboga, Oscar Alberto; Trans-complementation of polyhedrin by a stably transformed Sf9 insect cell line allows occ- baculovirus occlusion and larval per os infectivity; Elsevier Science; Journal of Biotechnology; 145; 2; 1-2010; 199-205
0168-1656
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/10.1016/j.jbiotec.2009.10.015
info:eu-repo/semantics/altIdentifier/url/https://www.sciencedirect.com/science/article/pii/S0168165609004842
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
application/pdf
dc.publisher.none.fl_str_mv Elsevier Science
publisher.none.fl_str_mv Elsevier Science
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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score 13.13397

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