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.
Instituto de Biotecnología
Fil: Lopez, Maria Gabriela. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Alfonso, Victoria. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Carrillo, Elisa Cristina. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Taboga, Oscar Alberto. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fuente
Journal of Biotechnology 145 (2) : 199-205 (January 2010)
Materia
Biotecnología
Insecta
Baculovirus
Genética
Biotechnology
Genetics
Polihedrina
Cuerpos de Oclusión
Polyhedrin
Occlusion Bodies
Nivel de accesibilidad
acceso restringido
Condiciones de uso
Repositorio
INTA Digital (INTA)
Institución
Instituto Nacional de Tecnología Agropecuaria
OAI Identificador
oai:localhost:20.500.12123/5110
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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 AlbertoBiotecnologíaInsectaBaculovirusGenéticaBiotechnologyGeneticsPolihedrinaCuerpos de OclusiónPolyhedrinOcclusion BodiesOral 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.Instituto de BiotecnologíaFil: Lopez, Maria Gabriela. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Alfonso, Victoria. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Carrillo, Elisa Cristina. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Taboga, Oscar Alberto. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaElsevier2019-05-14T13:45:07Z2019-05-14T13:45:07Z2010-01-15info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfhttps://www.sciencedirect.com/science/article/pii/S0168165609004842http://hdl.handle.net/20.500.12123/51100168-1656https://doi.org/10.1016/j.jbiotec.2009.10.015Journal of Biotechnology 145 (2) : 199-205 (January 2010)reponame:INTA Digital (INTA)instname:Instituto Nacional de Tecnología Agropecuariaenginfo:eu-repo/semantics/restrictedAccess2025-10-16T09:29:31Zoai:localhost:20.500.12123/5110instacron:INTAInstitucionalhttp://repositorio.inta.gob.ar/Organismo científico-tecnológicoNo correspondehttp://repositorio.inta.gob.ar/oai/requesttripaldi.nicolas@inta.gob.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:l2025-10-16 09:29:32.203INTA Digital (INTA) - Instituto Nacional de Tecnología Agropecuariafalse
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
Biotecnología
Insecta
Baculovirus
Genética
Biotechnology
Genetics
Polihedrina
Cuerpos de Oclusión
Polyhedrin
Occlusion Bodies
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 Biotecnología
Insecta
Baculovirus
Genética
Biotechnology
Genetics
Polihedrina
Cuerpos de Oclusión
Polyhedrin
Occlusion Bodies
topic Biotecnología
Insecta
Baculovirus
Genética
Biotechnology
Genetics
Polihedrina
Cuerpos de Oclusión
Polyhedrin
Occlusion Bodies
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.
Instituto de Biotecnología
Fil: Lopez, Maria Gabriela. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Alfonso, Victoria. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Carrillo, Elisa Cristina. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Taboga, Oscar Alberto. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; 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.
publishDate 2010
dc.date.none.fl_str_mv 2010-01-15
2019-05-14T13:45:07Z
2019-05-14T13:45:07Z
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 https://www.sciencedirect.com/science/article/pii/S0168165609004842
http://hdl.handle.net/20.500.12123/5110
0168-1656
https://doi.org/10.1016/j.jbiotec.2009.10.015
url https://www.sciencedirect.com/science/article/pii/S0168165609004842
http://hdl.handle.net/20.500.12123/5110
https://doi.org/10.1016/j.jbiotec.2009.10.015
identifier_str_mv 0168-1656
dc.language.none.fl_str_mv eng
language eng
dc.rights.none.fl_str_mv info:eu-repo/semantics/restrictedAccess
eu_rights_str_mv restrictedAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Elsevier
publisher.none.fl_str_mv Elsevier
dc.source.none.fl_str_mv Journal of Biotechnology 145 (2) : 199-205 (January 2010)
reponame:INTA Digital (INTA)
instname:Instituto Nacional de Tecnología Agropecuaria
reponame_str INTA Digital (INTA)
collection INTA Digital (INTA)
instname_str Instituto Nacional de Tecnología Agropecuaria
repository.name.fl_str_mv INTA Digital (INTA) - Instituto Nacional de Tecnología Agropecuaria
repository.mail.fl_str_mv tripaldi.nicolas@inta.gob.ar
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score 12.712165

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