Generation of a Recombinant Anticarsia Gemmatalis Multicapsid Nucleopolyhedrovirus Expressing a Foreign Gene under the Control of a Very Late Promoter
- Autores
- Arana, Eloisa Irene; Albariño, César G.; O'Reilly, David; Ghiringhelli, Daniel; Romanowski, Víctor
- Año de publicación
- 2001
- Idioma
- español castellano
- Tipo de recurso
- artículo
- Estado
- versión publicada
- Descripción
- A recombinant Anticarsia gemmatalis multicapsid nucleopolyhedrovirus (AgMNPV) expressing β-galactosidase under the control of the polyhedrin promoter was generated in our laboratory. To this end, we cloned the AgMNPV-2D genomic DNA fragment containing the polh gene and subcloned and sequenced the polyhedrin gene and its flanking regions. Based on this sequence information, sets of primers were designed to amplify the flanking regions by PCR, including appropriate restriction sites. The transfer vector (pAgPHZ) was constructed by the consecutive cloning of these PCR fragments flanking the Escherichia coli LacZ gene, in place of the polh gene. pAgPHZ was used for cotransfection of UFL-AG-286 insect cells with AgMNPV-2D DNA and the required recombinant, generated by homologous recombination with the polh locus, was identified by its polh⁻/LacZ⁺ plaque phenotype. Its genome structure was confirmed by PCR, restriction digestion and Southern blot analyses. The kinetics and levels of expression of β-galactosidase in UFL-AG-286 cells infected with the recombinant were tested by SDS-PAGE and enzymatic activity assays.
Instituto de Biotecnología y Biología Molecular - Materia
-
Ciencias Exactas
Biología
baculovirus
Anticarsia gemmatalis nucleopolyhedrovirus
AgMNPV
recombinant baculovirus
β-galactosidase
polyhedrin - Nivel de accesibilidad
- acceso abierto
- Condiciones de uso
- http://creativecommons.org/licenses/by/4.0/
- Repositorio
.jpg)
- Institución
- Universidad Nacional de La Plata
- OAI Identificador
- oai:sedici.unlp.edu.ar:10915/140153
Ver los metadatos del registro completo
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Generation of a Recombinant Anticarsia Gemmatalis Multicapsid Nucleopolyhedrovirus Expressing a Foreign Gene under the Control of a Very Late PromoterArana, Eloisa IreneAlbariño, César G.O'Reilly, DavidGhiringhelli, DanielRomanowski, VíctorCiencias ExactasBiologíabaculovirusAnticarsia gemmatalis nucleopolyhedrovirusAgMNPVrecombinant baculovirusβ-galactosidasepolyhedrinA recombinant <i>Anticarsia gemmatalis</i> multicapsid nucleopolyhedrovirus (AgMNPV) expressing β-galactosidase under the control of the polyhedrin promoter was generated in our laboratory. To this end, we cloned the AgMNPV-2D genomic DNA fragment containing the polh gene and subcloned and sequenced the polyhedrin gene and its flanking regions. Based on this sequence information, sets of primers were designed to amplify the flanking regions by PCR, including appropriate restriction sites. The transfer vector (pAgPHZ) was constructed by the consecutive cloning of these PCR fragments flanking the <i>Escherichia coli LacZ</i> gene, in place of the polh gene. pAgPHZ was used for cotransfection of UFL-AG-286 insect cells with AgMNPV-2D DNA and the required recombinant, generated by homologous recombination with the polh locus, was identified by its polh⁻/LacZ⁺ plaque phenotype. Its genome structure was confirmed by PCR, restriction digestion and Southern blot analyses. The kinetics and levels of expression of β-galactosidase in UFL-AG-286 cells infected with the recombinant were tested by SDS-PAGE and enzymatic activity assays.Instituto de Biotecnología y Biología Molecular2001-06info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionArticulohttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdf363-372http://sedici.unlp.edu.ar/handle/10915/140153spainfo:eu-repo/semantics/altIdentifier/issn/0920-8569info:eu-repo/semantics/altIdentifier/issn/1572-994Xinfo:eu-repo/semantics/altIdentifier/doi/10.1023/a:1011186828109info:eu-repo/semantics/altIdentifier/pmid/11450955info: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:UNLP2025-09-03T11:04:13Zoai:sedici.unlp.edu.ar:10915/140153Institucionalhttp://sedici.unlp.edu.ar/Universidad públicaNo correspondehttp://sedici.unlp.edu.ar/oai/snrdalira@sedici.unlp.edu.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:13292025-09-03 11:04:13.687SEDICI (UNLP) - Universidad Nacional de La Platafalse |
| dc.title.none.fl_str_mv |
Generation of a Recombinant Anticarsia Gemmatalis Multicapsid Nucleopolyhedrovirus Expressing a Foreign Gene under the Control of a Very Late Promoter |
| title |
Generation of a Recombinant Anticarsia Gemmatalis Multicapsid Nucleopolyhedrovirus Expressing a Foreign Gene under the Control of a Very Late Promoter |
| spellingShingle |
Generation of a Recombinant Anticarsia Gemmatalis Multicapsid Nucleopolyhedrovirus Expressing a Foreign Gene under the Control of a Very Late Promoter Arana, Eloisa Irene Ciencias Exactas Biología baculovirus Anticarsia gemmatalis nucleopolyhedrovirus AgMNPV recombinant baculovirus β-galactosidase polyhedrin |
| title_short |
Generation of a Recombinant Anticarsia Gemmatalis Multicapsid Nucleopolyhedrovirus Expressing a Foreign Gene under the Control of a Very Late Promoter |
| title_full |
Generation of a Recombinant Anticarsia Gemmatalis Multicapsid Nucleopolyhedrovirus Expressing a Foreign Gene under the Control of a Very Late Promoter |
| title_fullStr |
Generation of a Recombinant Anticarsia Gemmatalis Multicapsid Nucleopolyhedrovirus Expressing a Foreign Gene under the Control of a Very Late Promoter |
| title_full_unstemmed |
Generation of a Recombinant Anticarsia Gemmatalis Multicapsid Nucleopolyhedrovirus Expressing a Foreign Gene under the Control of a Very Late Promoter |
| title_sort |
Generation of a Recombinant Anticarsia Gemmatalis Multicapsid Nucleopolyhedrovirus Expressing a Foreign Gene under the Control of a Very Late Promoter |
| dc.creator.none.fl_str_mv |
Arana, Eloisa Irene Albariño, César G. O'Reilly, David Ghiringhelli, Daniel Romanowski, Víctor |
| author |
Arana, Eloisa Irene |
| author_facet |
Arana, Eloisa Irene Albariño, César G. O'Reilly, David Ghiringhelli, Daniel Romanowski, Víctor |
| author_role |
author |
| author2 |
Albariño, César G. O'Reilly, David Ghiringhelli, Daniel Romanowski, Víctor |
| author2_role |
author author author author |
| dc.subject.none.fl_str_mv |
Ciencias Exactas Biología baculovirus Anticarsia gemmatalis nucleopolyhedrovirus AgMNPV recombinant baculovirus β-galactosidase polyhedrin |
| topic |
Ciencias Exactas Biología baculovirus Anticarsia gemmatalis nucleopolyhedrovirus AgMNPV recombinant baculovirus β-galactosidase polyhedrin |
| dc.description.none.fl_txt_mv |
A recombinant <i>Anticarsia gemmatalis</i> multicapsid nucleopolyhedrovirus (AgMNPV) expressing β-galactosidase under the control of the polyhedrin promoter was generated in our laboratory. To this end, we cloned the AgMNPV-2D genomic DNA fragment containing the polh gene and subcloned and sequenced the polyhedrin gene and its flanking regions. Based on this sequence information, sets of primers were designed to amplify the flanking regions by PCR, including appropriate restriction sites. The transfer vector (pAgPHZ) was constructed by the consecutive cloning of these PCR fragments flanking the <i>Escherichia coli LacZ</i> gene, in place of the polh gene. pAgPHZ was used for cotransfection of UFL-AG-286 insect cells with AgMNPV-2D DNA and the required recombinant, generated by homologous recombination with the polh locus, was identified by its polh⁻/LacZ⁺ plaque phenotype. Its genome structure was confirmed by PCR, restriction digestion and Southern blot analyses. The kinetics and levels of expression of β-galactosidase in UFL-AG-286 cells infected with the recombinant were tested by SDS-PAGE and enzymatic activity assays. Instituto de Biotecnología y Biología Molecular |
| description |
A recombinant <i>Anticarsia gemmatalis</i> multicapsid nucleopolyhedrovirus (AgMNPV) expressing β-galactosidase under the control of the polyhedrin promoter was generated in our laboratory. To this end, we cloned the AgMNPV-2D genomic DNA fragment containing the polh gene and subcloned and sequenced the polyhedrin gene and its flanking regions. Based on this sequence information, sets of primers were designed to amplify the flanking regions by PCR, including appropriate restriction sites. The transfer vector (pAgPHZ) was constructed by the consecutive cloning of these PCR fragments flanking the <i>Escherichia coli LacZ</i> gene, in place of the polh gene. pAgPHZ was used for cotransfection of UFL-AG-286 insect cells with AgMNPV-2D DNA and the required recombinant, generated by homologous recombination with the polh locus, was identified by its polh⁻/LacZ⁺ plaque phenotype. Its genome structure was confirmed by PCR, restriction digestion and Southern blot analyses. The kinetics and levels of expression of β-galactosidase in UFL-AG-286 cells infected with the recombinant were tested by SDS-PAGE and enzymatic activity assays. |
| publishDate |
2001 |
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2001-06 |
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info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion Articulo http://purl.org/coar/resource_type/c_6501 info:ar-repo/semantics/articulo |
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http://sedici.unlp.edu.ar/handle/10915/140153 |
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