Construction of a recombinant Anticarsia gemmatalis nucleopolyhedrovirus (AgMNPV-2D) harbouring the beta-galactosidase gene

Construction of a recombinant Anticarsia gemmatalis nucleopolyhedrovirus (AgMNPV-2D) harbouring the beta-galactosidase gene

Autor Ribeiro, B. M. Google Scholar
Gatti, CDC Google Scholar
Costa, M. H. Google Scholar
Moscardi, F. Google Scholar
Maruniak, J. E. Google Scholar
Possee, R. D. Google Scholar
Zanotto, PMA Google Scholar
Instituição Universidade Federal de São Paulo (UNIFESP)
Universidade de Brasília (UnB)
Empresa Brasileira de Pesquisa Agropecuária (EMBRAPA)
NERC
Univ Florida
Resumo We have constructed a transfer vector (pAgGal) containing the beta -galactosidase gene under control of the Escherichia coli gpt and AgMNPV polyhedrin (polh) promoters. the transfer vector was cotransfected with wild type Anticarsia gemmatalis nucleopolyhedrovirus (AgMNPV) DNA into A. gemmatalis (UFL-AG-286) cells and a recombinant baculovirus (vAgGalA2) was isolated. the beta -galactosidase gene insertion was checked by polymerase chain reaction (PCR) using DNA from AgMNPV and vAgGalA2 and primers specific for regions upstream and downstream of the polh gene. Insect cells (UFL-AG-286) were infected with the recombinant vAgGalA2 and wild type AgMNPV viruses and the production of the heterologous protein analyzed by SDS-PAGE and Pulse-Chase. beta -galactosidase was expressed at high levels late on infection as expected for a gene under the control of the polh promoter. the highly expressed beta -galactosidase protein was also shown to be biologically active by a beta -galactosidase assay.
Idioma Inglês
Data de publicação 2001-01-01
Publicado em Archives of Virology. Vienna: Springer-verlag Wien, v. 146, n. 7, p. 1355-1367, 2001.
ISSN 0304-8608 (Sherpa/Romeo, fator de impacto)
Publicador Springer
Extensão 1355-1367
Fonte http://dx.doi.org/10.1007/s007050170096
Direito de acesso Acesso restrito
Tipo Artigo
Web of Science WOS:000170368200009
Endereço permanente http://repositorio.unifesp.br/handle/11600/26444

Exibir registro completo




Arquivo

Arquivo Tamanho Formato Visualização

Não existem arquivos associados a este item.

Este item está nas seguintes coleções

Buscar


Navegar

Minha conta