Generation of a Recombinant Anticarsia Gemmatalis Multicapsid Nucleopolyhedrovirus Expressing a Foreign Gene under the Control of a Very Late Promoter

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 a...

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Detalles Bibliográficos
Autores principales:	Arana, Eloisa Irene, Albariño, César G., O'Reilly, David, Ghiringhelli, Daniel, Romanowski, Víctor
Formato:	Articulo
Lenguaje:	Español
Publicado:	2001
Materias:	Ciencias Exactas Biología baculovirus Anticarsia gemmatalis nucleopolyhedrovirus AgMNPV recombinant baculovirus β-galactosidase polyhedrin
Acceso en línea:	http://sedici.unlp.edu.ar/handle/10915/140153
Aporte de:	SEDICI (UNLP) de Universidad Nacional de La Plata

id	I19-R120-10915-140153
record_format	dspace
institution	Universidad Nacional de La Plata
institution_str	I-19
repository_str	R-120
collection	SEDICI (UNLP)
language	Español
topic	Ciencias Exactas Biología baculovirus Anticarsia gemmatalis nucleopolyhedrovirus AgMNPV recombinant baculovirus β-galactosidase polyhedrin
spellingShingle	Ciencias Exactas Biología baculovirus Anticarsia gemmatalis nucleopolyhedrovirus AgMNPV recombinant baculovirus β-galactosidase polyhedrin Arana, Eloisa Irene Albariño, César G. O'Reilly, David Ghiringhelli, Daniel Romanowski, Víctor Generation of a Recombinant Anticarsia Gemmatalis Multicapsid Nucleopolyhedrovirus Expressing a Foreign Gene under the Control of a Very Late Promoter
topic_facet	Ciencias Exactas Biología baculovirus Anticarsia gemmatalis nucleopolyhedrovirus AgMNPV recombinant baculovirus β-galactosidase polyhedrin
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.
format	Articulo Articulo
author	Arana, Eloisa Irene Albariño, César G. O'Reilly, David Ghiringhelli, Daniel Romanowski, Víctor
author_facet	Arana, Eloisa Irene Albariño, César G. O'Reilly, David Ghiringhelli, Daniel Romanowski, Víctor
author_sort	Arana, Eloisa Irene
title	Generation of a Recombinant Anticarsia Gemmatalis Multicapsid Nucleopolyhedrovirus Expressing a Foreign Gene under the Control of a Very Late Promoter
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
publishDate	2001
url	http://sedici.unlp.edu.ar/handle/10915/140153
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Generation of a Recombinant Anticarsia Gemmatalis Multicapsid Nucleopolyhedrovirus Expressing a Foreign Gene under the Control of a Very Late Promoter

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